1. The shorthorn sculpin antifreeze protein preferentially binds along the [1 2 2] direction of the (2 1_ 0) face of ice.
2. Residues such as K9, K31, R12, and K23 play crucial roles in binding to ice.
3. Compared with the winter flounder antifreeze protein, the degree of fit between the shorthorn sculpin antifreeze protein and the ice surface is important.
AFP011007
General Information
| Protein Name | Myoxocephalus scorpius SS-8 AFP |
| Species | Myoxocephalus scorpius (Shorthorn sculpin) (Cottus scorpius) |
| Sequence Length | 42 |
| Sequence | |
| Structure | AF2 predicted Structure: AFP011007 |
| Solvent Accessible Surface Area | Total SASA | 2639.32 Ų | Polar SASA | 1020.33 Ų | Apolar SASA | 3659.65 Ų |
AFP011007000
| Mutation | Wild Type |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | wild-type SS-8 AFP |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | wild-type SS-8 AFP |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Ice Binding Sites
| PMID | 8804585 |
| DOI | 10.1016/S0006-3495(96)79204-4 |
| Protein Name | shorthorn sculpin AFP |
| IBS | The binding sites involve K9, K31, R12, and K23. |
Ice Binding Sites
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | wild-type SS-8 AFP |
| IBS | The binding site is the Ala-rich surface, and specific Ala residues such as Ala10, Ala14, Ala21, Ala25, Ala32, and Ala36 have surface complementarity with the grooves of the ice surface. |
Ice plane
| PMID | 8804585 |
| DOI | 10.1016/S0006-3495(96)79204-4 |
| Protein Name | shorthorn sculpin AFP |
| Ice plane | The antifreeze protein binds to the (2 1_ 0) plane of ice along the [1 2 2] direction. |
Ice plane
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | wild-type SS-8 AFP |
| Ice plane | The antifreeze protein binds to the (1 1_ 2 0) plane. |
Brief description
|
PMID: 8804585
DOI: 10.1016/S0006-3495(96)79204-4 |
|
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007001
| Mutation | 35_42del,34_35insAAAAAATAR,1_8del,0_1insDTASDAAA |
| Sequence |
Ice crystal morphology
| PMID | 11710110 |
| DOI | 10.1021/bm000004w |
| Protein Name | winter flounder AFP(43 mer) |
| Ice crystal morphology | Fig.3.A,C Morphology of ice crystals grown from 100 mg/mL solution of the 43-mer within the thermal hysteresis, i.e., before the nonequilibrium freezing point was reached (magnification 400x). Morphology of ice crystals grown from a 250 mg/mL solution of the 43-mer when the nonequilibrium freezing point is reached. Ice crystals grow in the form of needles along the c axis of ice (magnification 400×). |
Thermal Hysteresis
| PMID | 11710110 |
| DOI | 10.1021/bm000004w |
| Protein Name | winter flounder AFP(43 mer) |
| Thermal Hysteresis | Fig.4 Nonequilibrium freezing point depression of the 43-mer vs concentration. Note that there was no nonequilibrium antifreeze activity observed below 50 mg/mL. |
Ice plane
| PMID | 11710110 |
| DOI | 10.1021/bm000004w |
| Protein Name | winter flounder AFP(43 mer) |
| Ice plane | The antifreeze protein binds to the (2 1_ 0) secondary prism planes of ice along the [1 2 2] direction. |
Brief description
|
PMID: 11710110
DOI: 10.1021/bm000004w |
1. A lysine-alanine-rich 43-mer polypeptide was successfully de novo designed, which exhibits nonequilibrium freezing point depression.
2. Similar to the shorthorn sculpin AFP, the 43-mer polypeptide binds to the (2 1_ 0) secondary prism planes of ice along the [1 2 2] direction. 3. The artificially designed 32-mer and 27-mer polypeptides have no antifreeze activity at the tested concentrations due to structural problems such as low helical content. |
AFP011007002
| Mutation | 35_42del,34_35insAA,1_8del,0_1insAAAA |
| Sequence |
Thermal Hysteresis
| PMID | 11710110 |
| DOI | 10.1021/bm000004w |
| Protein Name | winter flounder AFP(32-mer) |
| Thermal Hysteresis | 'As indicated before, the 27-mer and the 32-mer were inactive at the concentrations used for the study (~10 mg/mL), showing no ice morphology modifications.' |
Brief description
|
PMID: 11710110
DOI: 10.1021/bm000004w |
1. A lysine-alanine-rich 43-mer polypeptide was successfully de novo designed, which exhibits nonequilibrium freezing point depression.
2. Similar to the shorthorn sculpin AFP, the 43-mer polypeptide binds to the (2 1_ 0) secondary prism planes of ice along the [1 2 2] direction. 3. The artificially designed 32-mer and 27-mer polypeptides have no antifreeze activity at the tested concentrations due to structural problems such as low helical content. |
AFP011007003
| Mutation | 34_42del,1_8del,0_1insAQ |
| Sequence |
Thermal Hysteresis
| PMID | 11710110 |
| DOI | 10.1021/bm000004w |
| Protein Name | winter flounder AFP(27-mer) |
| Thermal Hysteresis | 'As indicated before, the 27-mer and the 32-mer were inactive at the concentrations used for the study (~10 mg/mL), showing no ice morphology modifications.' |
Brief description
|
PMID: 11710110
DOI: 10.1021/bm000004w |
1. A lysine-alanine-rich 43-mer polypeptide was successfully de novo designed, which exhibits nonequilibrium freezing point depression.
2. Similar to the shorthorn sculpin AFP, the 43-mer polypeptide binds to the (2 1_ 0) secondary prism planes of ice along the [1 2 2] direction. 3. The artificially designed 32-mer and 27-mer polypeptides have no antifreeze activity at the tested concentrations due to structural problems such as low helical content. |
AFP011007004
| Mutation | 41_42del |
| Sequence |
Ice plane
| PMID | 2009357 |
| DOI | 10.1016/S0006-3495(91)82234-2 |
| Protein Name | Short-hom Sculpin AFP |
| Ice plane | It binds to the six equivalent (2 1_ 1_ 0) secondary prism planes. |
Brief description
|
PMID: 2009357
DOI: 10.1016/S0006-3495(91)82234-2 |
1. The peptide from short-horn sculpin adsorbs on the (2 1_ 1_ 0) secondary prism planes.
2. The alignment directions of the three antifreeze peptides on the adsorption planes are probably all [0 1 1_ 2], which is deduced from the elongation directions of the etched regions. |
AFP011007005
| Mutation | A16K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A16K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A16K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007006
| Mutation | A17K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A17K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A17K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007007
| Mutation | A19K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A19K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A19K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007008
| Mutation | A21K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A21K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A21K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007009
| Mutation | A22K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A22K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A22K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |
AFP011007010
| Mutation | A25K |
| Sequence |
Ice crystal morphology
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A25K) |
| Ice crystal morphology | Fig.5 Ice crystal morphology in the presence of wild-type SS-8 and variants. (A) Ice crystals formed in the presence of 1.0 mg/mL WT,A16K,A19K,A22K,or A25K in 0.1 M NH4HCO3. The samples were undercooled to 0.09℃ over 75 sec,after which image collection timing began from time zero. SS-8 variants that generated wild-type ice crystal morphology included A16K,A19K,and A22K. The inactive variant A25K ice crystal grew slowly over the 10-min interval of measurement,and generated an elongated hexagonal bipyramid. (B) Ice crystals of the highly inactive samples were formed in the presence of 1.0 mg/mL A17K or A21K in 0.1 M NH4HCO3, and image collection began immediately after ice crystal stabilization at 0.02℃ of undercooling. |
Thermal Hysteresis
| PMID | 11714925 |
| DOI | 10.1110/ps.ps.26501 |
| Protein Name | shorthorn sculpin SS-8 AFP(A25K) |
| Thermal Hysteresis | Fig.4 Thermal hysteresis activity as a function of antifreeze protein concentration (mg/mL). Active antifreeze variants are: WT (●),A16K (○), A19K (▼),A22K (▽). Inactive antifreeze variants are: A17K (■),A21K (□),and A25K (◆). |
Brief description
|
PMID: 11714925
DOI: 10.1110/ps.ps.26501 |
1. The ice-binding surface of the shorthorn sculpin antifreeze protein SS-8 is the Ala-rich surface, not the Lys-rich hydrophilic surface.
2. Single Ala to Lys substitution experiments on SS-8 showed that substitutions on the hydrophobic (Ala-rich) surface (such as A17K, A21K, A25K) abolished antifreeze activity, while those on the hydrophilic surface (such as A16K, A19K, A22K) had no obvious effect. |