Extremely elastic spider silk protein and DNA coding therefor
AssigneeUniversity of Wyoming
InventorsRandolph V. Lewis, Cheryl Y. Hayashi
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Abstract
Cloned DNA encoding spider flagelliform silk proteins described. The translated amino acid sequence of the cloned cDNA shows that the flagelliform silk protein is composed largely of repeated motifs. The dominant motif of this protein is Gly-Pro-Gly-Gly-X, which can appear up to 63 times in tandem arrays. This motif likely forms Pro.sup.2 -Gly.sup.3 type II .beta.-turns. The resulting series of concatenated .beta.-turns are thought to form a helix (.beta.-spiral). The present inventors propose that this spring-like .beta.-spiral is the basis of the elasticity of silk.
Claims71
An isolated and purified flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-X).sub.n
- wherein n is an integer of at least 4, wherein X is an amino acid, wherein 75 to 95% of X are selected from the group consisting of Ala, Ser, Tyr, and Val.
An isolated and purified flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ala-Gly-Pro-Gly-Gly-Ala).sub.n
- wherein n is an integer of at least 2.
An isolated and purified flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ser-Gly-Pro-Gly-Gly-Tyr).sub.n
- wherein n is an integer of at least 2.
An isolated and purified flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Tyr-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Val-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-X).sub.n
- wherein n is an integer of at least 4, wherein X is an amino acid, wherein 75 to 95% of X are selected from the group consisting of Ala, Ser, Tyr, and Val.
An isolated and purified protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ala-Gly-Pro-Gly-Gly-Ala).sub.n
- wherein n is an integer of at least 2.
An isolated and purified protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ser-Gly-Pro-Gly-Gly-Tyr).sub.n
- wherein n is an integer of at least 2.
An isolated and purified protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Tyr-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Val-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified protein having the formula:
- [(Gly-Pro-Gly-Gly-X.sub.1).sub.n (Gly-Gly-X.sub.2).sub.m S (Gly-Gly-X.sub.2).sub.m ].sub.p
- wherein n is an integer between 43 and 63, wherein X.sub.1 is an amino acid, wherein 75 to 95% of X.sub.1 are selected from the group consisting of Ala, Ser, Tyr, and Val;
- wherein m is an integer between 3 and 12, wherein X.sub.2 is an amino acid, wherein 80-99% of X.sub.2 are selected from the group consisting of Ala, Ser, and Thr;
- wherein S is an amino acid spacer sequence comprising 23 to 33 amino acids;
- and p is an integer greater than 1.
An isolated and purified nucleic acid molecule encoding a flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-X).sub.n
- wherein n is an integer of at least 4, wherein X is an amino acid, wherein 75 to 95% of X are selected from the group consisting of Ala, Ser, Tyr, and Val.
An isolated and purified nucleic acid molecule encoding a flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ala-Gly-Pro-Gly-Gly-Ala).sub.n
- wherein n is an integer of at least 2.
An isolated and purified nucleic acid molecule encoding a flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Ser-Gly-Pro-Gly-Gly-Tyr).sub.n
- wherein n is an integer of at least 2.
An isolated and purified nucleic acid molecule encoding a flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Tyr-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified nucleic acid molecule encoding a flagelliform gland silk fibroin protein originating from a flagelliform gland of an orb-web spinning spider, said silk fibroin protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-Val-Gly-Pro-Gly-Gly-Ser).sub.n
- wherein n is an integer of at least 2.
An isolated and purified nucleic acid molecule encoding protein comprising a glycine and proline rich repetitive .beta.s region that forms a .beta.-spiral, said .beta.s region represented by the following formula:
- (Gly-Pro-Gly-Gly-X).sub.n
- wherein n is an integer of at least 4, wherein X is an amino acid, wherein 75 to 95% of X are selected from the group consisting of Ala, Ser, Tyr, and Val.
An isolated and purified nucleic acid molecule encoding a protein having the formula:
- [(Gly-Pro-Gly-Gly-X.sub.1).sub.n (Gly-Gly-X.sub.2).sub.m S (Gly-Gly-X.sub.2).sub.m ].sub.p
- wherein n is an integer between 43 and 63, wherein X.sub.1 is an amino acid, wherein 75 to 95% of X.sub.1 are selected from the group consisting of Ala, Ser, Tyr, and Val, wherein m is an integer between 3 and 12, wherein X.sub.2 is an amino acid, wherein 80-99% of X.sub.2 are selected from the group consisting of Ala, Ser, and Thr, wherein S is an amino acid spacer sequence comprising 23 to 33 amino acids, and wherein p is an integer greater than 1.
Dependencies are inferred from the claim text.