1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth.
3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears.
AFP012000
General Information
| Protein Name | Antifreeze protein |
| Gene Name in Uniprot | AFP type II |
| UniProt ID | Q91992 |
| Nucleotide Sequence ID in NCBI | L14722 / S65819.1 |
| Protein Sequence ID in NCBI | AAA49200.1 / AAB28093.1 |
| Species | Clupea harengus (Atlantic herring) |
| Sequence Length | 130 |
| Sequence | |
| Structure | PDB ID: 2py2 |
| Solvent Accessible Surface Area | Total SASA | 21217.21 Ų | Polar SASA | 13432.20 Ų | Apolar SASA | 34649.42 Ų |
AFP012000000
| Mutation | Wild Type |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP WT-6H |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.A Ice crystal morphology of hAFP and its CaCR mutants. The samples were (A) buffer alone and wild-type hAFP WT-6H. The protein concentrations used for each sample are indicated. |
Ice crystal morphology
| PMID | 9521729 |
| DOI | 10.1021/bi972503w |
| Protein Name | hAFP(herring AFP) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphologies in the AFP solutions used to measure thermal hysteresis. Protein concentrations are 0.10, 0.5, 0.75, and 1.5 mg/mL in panels 1-4, respectively. A protein-free control is also shown. The magnification used was 400X. |
Ice crystal morphology
| PMID | 8663288 |
| DOI | 10.1074/jbc.271.28.16627 |
| Protein Name | hAFP |
| Ice crystal morphology | Fig.6 Antifreeze activity of hAFP in the presence of metals. Ice crystals were grown in solutions of 4 mg/ml AFP. Crystals are shown with the a and c axes indicated. An exception is the crystal grown in the absence of added ion that has the a axis parallel to the page and the c axis normal to the page. Added metal ions are indicated. Crystals were videotaped at temperatures below the melting point. Individual frames were then transferred into image files. |
Ice crystal morphology
| PMID | 16887111 |
| DOI | 10.1016/j.cryobiol.2006.06.006 |
| Protein Name | Recombinant type II AFP (herring) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.2.(3) Ice crystal bursts in the presence of moderately active Wsh AFPs. Images of ice crystals seen in frames A-D were taken at 0.4 s intervals during the burst. Sequences 1-6 were obtained with type I AFP from winter Xounder plasma, shorthorn sculpin plasma, recombinant type II AFP (herring), recombinant type II (sea raven), recombinant type III AFP and low molecular weight AFGPs from Gadus ogac, respectively. The AFP concentrations and thermal hysteresis activities for each AFP are given in Table 1. Each AFP was assayed in 100 mM ammonium bicarbonate (pH 7.9). The herring type II AFP sample also contained 10 mM CaCl2. The direction of the c-axis is shown in frame A by the white arrow. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP WT-6H |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.3 Thermal hysteresis activity of hAFP and its CaCR mutants Q92A, D94A, N113A, and N113D. (A) Measurements of concentration-dependent thermal hysteresis of wild-type hAFP WT6H and its mutants. (B) Relative thermal hysteresis of hAFP CaCR mutants (0.4 mM) compared with that of WT-6H. |
Thermal Hysteresis
| PMID | 9521729 |
| DOI | 10.1021/bi972503w |
| Protein Name | hAFP(herring AFP) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.4.A Antifreeze activity (thermal hysteresis) of solutions containing a series of AFP concentrations was measured as described under Experimental Procedures using a Clifton nanoliter osmometer. Values shown represent means ±SD of triplicate measurements done on single samples. The curve was fitted using the data pooled from the native and recombinant AFPs. |
Thermal Hysteresis
| PMID | 8663288 |
| DOI | 10.1074/jbc.271.28.16627 |
| Protein Name | hAFP |
| Thermal Hysteresis | Fig.5 Antifreeze activity of hAFP as a function of added metals. A, antifreeze activity of solutions containing 4 mg/ml AFP and a series of CaCl2 concentrations was measured. Antifreeze activity of AFP was also measured in the absence of metal ions and in the presence of 10 mM metal ions.(B) and 3 mM ruthenium red as described under “Experimental Procedures” (C). For B and C, the average activity in the presence of Ca21 was normalized to 100%. Each bar represents the mean 6 S.E. of three preparations. |
Thermal Hysteresis
| PMID | 16887111 |
| DOI | 10.1016/j.cryobiol.2006.06.006 |
| Protein Name | Recombinant type II AFP (herring) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Tab.1 Recombinant type II (herring). |
Thermal Hysteresis
| PMID | 11281719 |
| DOI | 10.1006/prep.2001.1395 |
| Protein Name | Native |
| Thermal Hysteresis | Fig.7 Antifreeze activities of WT and WT-6H. Concentration-dependent antifreeze activities were measured as described under Materials and Methods using a Clifton nanoliter osmometer. Valuesshown represent means of triplicate measure ments done on eachsingle sample. |
Thermal Hysteresis
| PMID | 11281719 |
| DOI | 10.1006/prep.2001.1395 |
| Protein Name | hAFP WT-6H |
| Thermal Hysteresis | Fig.7 Antifreeze activities of WT and WT-6H. Concentration-dependent antifreeze activities were measured as described under Materials and Methods using a Clifton nanoliter osmometer. Valuesshown represent means of triplicate measure ments done on eachsingle sample. |
Thermal Hysteresis
| PMID | 11281719 |
| DOI | 10.1006/prep.2001.1395 |
| Protein Name | hAFP WT |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7 Antifreeze activities of WT and WT-6H. Concentration-dependent antifreeze activities were measured as described under Materials and Methods using a Clifton nanoliter osmometer. Valuesshown represent means of triplicate measure ments done on eachsingle sample. |
Ice Binding Sites
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP WT-6H |
| Tag | Hexahistidine |
| IBS | The ice-binding site(IBS) is located at the Ca2²⁺-binding site (CaBS), and the loop region defined by residues E99 and D114 is crucial for ice binding. |
Ice Binding Sites
| PMID | 9521729 |
| DOI | 10.1021/bi972503w |
| Protein Name | hAFP(herring AFP) |
| Tag | Hexahistidine |
| IBS | The ice-binding site is located at the Ca²⁺-binding site, which corresponds to the carbohydrate-binding site of C-type lectins. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
|
|
PMID: 9521729
DOI: 10.1021/bi972503w |
1. The refolded rAFP binds Ca²⁺ and exhibits a Ca²⁺-dependent protease resistance as the native AFP does.
2. The ice-binding site of the Atlantic herring type II antifreeze protein corresponds to the carbohydrate-binding site of C-type lectins. 3. The ice-binding function of the antifreeze protein may have evolved from the carbohydrate-binding site of a pre-existing C-type lectin. |
|
PMID: 8663288
DOI: 10.1074/jbc.271.28.16627 |
1. The results show that hAFP requires a single Ca²⁺ binding site, where key residues E99 and D114 are essential for both calcium coordination and ice binding, enabling thermal hysteresis (TH) activity.
2. In the absence of Ca²⁺ hAFP loses its antifreeze function, highlighting the crucial role of metal ion binding in maintaining its structural stability and activity. |
|
PMID: 16887111
DOI: 10.1016/j.cryobiol.2006.06.006 |
1. The study reveals that hyperactive AFPs are significantly more effective at depressing the freezing point of solutions than moderately active fish AFPs.
2. The key distinction lies in their ability to prevent ice growth out of the basal planes, resulting in ice crystals that burst explosively perpendicular to the c-axis when cooled below the freezing temperature. 3. This mechanism is proposed to be the foundation for the hyperactivity of these proteins. |
|
PMID: 11281719
DOI: 10.1006/prep.2001.1395 |
1. Recombinant herring antifreeze protein was successfully expressed in Pichia pastoris and exhibited antifreeze activity comparable to the native protein.
2. The C-terminal histidine tag has a minimal effect on the antifreeze activity of herring antifreeze protein. 3. The antifreeze activity and conformation of antifreeze proteins are Ca²⁺-dependent, and recombinant antifreeze proteins also have Ca²⁺-binding ability. |
AFP012000001
| Mutation | Q92E,D94N |
| Sequence |
Ice crystal morphology
| PMID | 9521729 |
| DOI | 10.1021/bi972503w |
| Protein Name | hAFP(EPN,mutant) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphologies in the AFP solutions used to measure thermal hysteresis. Protein concentrations are 0.10, 0.5, 0.75, and 1.5 mg/mL in panels 1-4, respectively. A protein-free control is also shown. The magnification used was 400X. |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(EPN) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 9521729 |
| DOI | 10.1021/bi972503w |
| Protein Name | hAFP(EPN,mutant) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.4.A Antifreeze activity (thermal hysteresis) of solutions containing a series of AFP concentrations was measured as described under Experimental Procedures using a Clifton nanoliter osmometer. Values shown represent means ±SD of triplicate measurements done on single samples. The curve was fitted using the data pooled from the native and recombinant AFPs. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(EPN) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "It is noted that mutant EPN showed only a trace of thermal hysteresis, even at 0.96 mM protein concentration." |
Brief description
|
PMID: 9521729
DOI: 10.1021/bi972503w |
1. The ice-binding site of the Atlantic herring type II antifreeze protein corresponds to the carbohydrate-binding site of C-type lectins.
2. The mutant antifreeze protein can bind Ca²⁺ normally and fold correctly, but it loses its antifreeze activity. 3. The ice-binding function of the antifreeze protein may have evolved from the carbohydrate-binding site of a pre-existing C-type lectin. |
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000002
| Mutation | Q92A |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphology of hAFP and its CaCR mutants. The samples were (B) hAFP mutants with reduced thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.3 Thermal hysteresis activity of hAFP and its CaCR mutants Q92A, D94A, N113A, and N113D. (A) Measurements of concentration-dependent thermal hysteresis of wild-type hAFP WT6H and its mutants. (B) Relative thermal hysteresis of hAFP CaCR mutants (0.4 mM) compared with that of WT-6H. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000003
| Mutation | D94A |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphology of hAFP and its CaCR mutants. The samples were (B) hAFP mutants with reduced thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.3 Thermal hysteresis activity of hAFP and its CaCR mutants Q92A, D94A, N113A, and N113D. (A) Measurements of concentration-dependent thermal hysteresis of wild-type hAFP WT6H and its mutants. (B) Relative thermal hysteresis of hAFP CaCR mutants (0.4 mM) compared with that of WT-6H. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000004
| Mutation | N113A |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(N113A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphology of hAFP and its CaCR mutants. The samples were (B) hAFP mutants with reduced thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(N113A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.3 Thermal hysteresis activity of hAFP and its CaCR mutants Q92A, D94A, N113A, and N113D. (A) Measurements of concentration-dependent thermal hysteresis of wild-type hAFP WT6H and its mutants. (B) Relative thermal hysteresis of hAFP CaCR mutants (0.4 mM) compared with that of WT-6H. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000005
| Mutation | N113D |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(N113D) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.B Ice crystal morphology of hAFP and its CaCR mutants. The samples were (B) hAFP mutants with reduced thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(N113D) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.3 Thermal hysteresis activity of hAFP and its CaCR mutants Q92A, D94A, N113A, and N113D. (A) Measurements of concentration-dependent thermal hysteresis of wild-type hAFP WT6H and its mutants. (B) Relative thermal hysteresis of hAFP CaCR mutants (0.4 mM) compared with that of WT-6H. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000006
| Mutation | Q92E |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92E) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92E) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "... which retained the ability to modify the ice crystal with no detectable thermal hysteresis." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000007
| Mutation | Q92T |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92T) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92T) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "... which retained the ability to modify the ice crystal with no detectable thermal hysteresis." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000008
| Mutation | D94N |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94N) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94N) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "... which retained the ability to modify the ice crystal with no detectable thermal hysteresis." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000009
| Mutation | E99A |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | E99A only modified the ice crystal with no detectable thermal hysteresis activity. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000010
| Mutation | E99D |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99D) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99D) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "... which retained the ability to modify the ice crystal with no detectable thermal hysteresis." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000011
| Mutation | E99Q |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99Q) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99Q) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "... which retained the ability to modify the ice crystal with no detectable thermal hysteresis." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000012
| Mutation | E99Q,N113D |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99Q/N113D) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.C Ice crystal morphology of hAFP and its CaCR mutants. The samples were (C) hAFP mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99Q/N113D) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Activity measurement for the double mutant E99Q/ N113D showed normal Calcium ion binding affinity with no detectable thermal hysteresis. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000013
| Mutation | Q92K |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92K) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(Q92K) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "The nonactive mutant Q92K … abolished the antifreeze activity completely." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000014
| Mutation | D94E |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94E) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94E) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "mutants with longer side chains, such as Q92K and D94E, … abolished the antifreeze activity completely." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000015
| Mutation | D94T |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D94T) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000016
| Mutation | E99H |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99H) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(E99H) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "E99H, which contained a bulky residue that might cause steric hindrance, completely lost its activity." |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000017
| Mutation | D114A |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D114A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D114A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | D114A was devoid of any thermal hysteresis. |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000018
| Mutation | D114E |
| Sequence |
Ice crystal morphology
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D114E) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.4.D Ice crystal morphology of hAFP and its CaCR mutants. The samples were (D) hAFP mutants that completely lost thermal hysteresis. The protein concentrations used for each sample are indicated. |
Thermal Hysteresis
| PMID | 15544325 |
| DOI | 10.1021/bi048485h |
| Protein Name | hAFP(D114E) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "Isoelectric alternates D94E, E99D, and D114E did not show higher antifreeze activity…" |
Brief description
|
PMID: 15544325
DOI: 10.1021/bi048485h |
1. The ice-binding site of herring type II antifreeze protein hAFP is located at the Ca²⁺-binding site.
2. Residues E99 and D114 are essential for the activity of hAFP to inhibit ice crystal growth. 3. Different mutations have different effects on the thermal hysteresis activity of hAFP. The thermal hysteresis activity of some mutants is significantly reduced or even disappears. |
AFP012000019
| Mutation | A90H |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A90H) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A90H) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.1 (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000020
| Mutation | A90S |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A90S) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A90S) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.1 (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000021
| Mutation | A91H |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A91H) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A91H) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.1 (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000022
| Mutation | A91T |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A91T) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (A91T) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.1 (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000023
| Mutation | T95A |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T95A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T95A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.2 T95A (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000024
| Mutation | T95I |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T95I) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T95I) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.2 T95A (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000025
| Mutation | G109D |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (G109D) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(2) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (G109D) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.2 T95A (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000026
| Mutation | Q103A |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (Q103A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (Q103A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.2 T95A (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000027
| Mutation | H121A |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (H121A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(3) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (H121A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | Fig.7.A.2 T95A (A) Concentration dependent thermal hysteresis activity of (1) WT-6H (●), mutants Ala90Ser (□), Ala90His (◼), Ala91His (▲), and Ala91Thr (△), and (2) WT-6H (●), mutants Thr95Ala (▲), Thr95Ile (△), Gln103Ala (□), Gly109Asp (○), and His121Ala (◼), was measured as described under "Materials and Methods" using a Clifton nanoliter osmometer. Values showed represent means of triplicate measurements done on each single sample. |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000028
| Mutation | T96I |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T96I) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(5) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T96I) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "In extreme cases, ice crystals of mutants Thr96Ile and Thr98Ala are circular plates similar to the mock control, indicating the complete loss of the ice-binding affinity" |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |
AFP012000029
| Mutation | T98A |
| Sequence |
Ice crystal morphology
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T98A) |
| Tag | Hexahistidine |
| Ice crystal morphology | Fig.7.B.(5) Ice crystal morphologies of hAFP and its mutants. Samples were: (1) buffer alone and wild-type hAFP; (2) the mutant that exhibited no effect on thermal hysteresis; (3) mutants that showed reduced thermal hysteresis activities; (4) mutants which retained the ability to modify the ice crystal with no detectable thermal hysteresis activities, (5) mutants that exhibited complete loss of antifreeze activity. The protein concentration used for each sample is also indicated. |
Thermal Hysteresis
| PMID | 17579720 |
| DOI | 10.1371/journal.pone.0000548 |
| Protein Name | hAFP (T98A) |
| Tag | Hexahistidine |
| Thermal Hysteresis | "In extreme cases, ice crystals of mutants Thr96Ile and Thr98Ala are circular plates similar to the mock control, indicating the complete loss of the ice-binding affinity" |
Brief description
|
PMID: 17579720
DOI: 10.1371/journal.pone.0000548 |
1. Determined the 1.7 Å crystal structure of herring type II AFP(hAFP), which resembles the C-type lectins and has unique ice-binding features.
2. Identified the ice-binding site of hAFP consisting of Thr96, Thr98, and Ca²⁺ coordinating residues Asp94 and Glu99, and the Ca²⁺ ion strengthens the interaction with ice. 3. Phylogenetic analysis showed that all type II AFPs evolved from a common ancestor and developed different ice-binding modes, originating from an ancestral duplication of a gene encoding a mannose or galactose-binding lectin. |