Polypeptide monomers, linearly extended and/or crosslinked forms thereof, and applications thereof

• US 5,574,134 A
• Filed: 02/19/1992
• Issued: 11/12/1996
• Est. Priority Date: 07/11/1989
• Status: Expired due to Fees

• Current Assignee: University of Delaware
• Original Assignee: University of Delaware
• Inventors: Waite, J. Herbert
• Primary Examiner(s): Seidleck, James J.
• Assistant Examiner(s): BOYKIN, TERRESSA M
• Application Number:
  US07/839,745
• Time in Patent Office: 1,728 Days
• Field of Search: 530/331, 530/328
• US Class Current: 530/328
• CPC Class Codes: C07K 14/43504 from invertebrates

Abstract

A water-resistant polyphenolic protein adhesive has been obtained from the Atlantic ribbed mussel, Geukensia (Modiolus) demissa. This protein provides a natural model for synthetic or bioengineered adhesives and is characterized by a Gly-Tyr-Lys or, more frequently, Gly-Dop-Lys "tail" fragment of a repeating octapeptide or nonapeptide unit. A complete octapeptide or nonapeptide unit (including both the 5- or 6-amino acid residue "head" as well as the 3-residue "tail") can be represented as follows:

[Gln]Glu].sub.x -Thr/Ala-Gly-Dop/Tyr-Y.sup.1 -Y.sup.2 -Gly-Tyr/Dop-Lys

where x is zero or one,

Y¹ is Val, Leu, Asp or Ser, and

Y² is Ala, Pro, Hyp, or Leu.

The tripeptide "tail" and/or the 5- or 6-residue "head" of the polypeptide is useful as a single unit (curable with difunctional crosslinkers or the like) or as a repeating unit.

Claims

1. An isolated and purified polypeptide containing at least one of the amino acid sequences set forth below:

(A) Gly-Dop-Lys

(B) Thr-Gly-Dop-Ser-Ala-Gly-Dop-Lys

(C) Gln-Thr-Gly-Dop-Val-Pro-Gly-Dop-Lys

(D) Gln-Thr-Gly-Dop-Asp-Pro-Gly-Tyr-Lys

(E) Gln-Thr-Gly-Dop-Leu-Pro-Gly-Dop-Lys

wherein the above three-letter symbols are defined as follows;

Gly is a glycine residue,

Dop is a 3,4-dihydroxyphenylalanine residue,

Lys is a lysine residue,

Gln is a glutamine residue,

Thr is a threonine residue,

Ser is a serine residue,

Ala is an alanine residue,

Val is a valine residue,

Pro is a proline residue,

Asp is an aspartic acid residue,

Tyr is a tyrosine residue and

Leu is a leucine residue.

2. A synthetic tripepride, octapeptide, or nonapeptide, wherein the synthetic tripeptide has the sequence (I)

View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11)

3. A synthetic homopolymer or random or block copolymer obtained by polymerizing, head/tail, up to about 50 tripeptides, octapeptides, or nonapeptides of the sequences (I), (II), and (III) of claim 2, whereby the total number of said sequences in the homopolymer or random or block copolymer does not exceed about 50.

4. A synthetic polymer consisting essentially of up to 50 head/tail polymerized tripeptides (I) of claim 2.

5. A synthetic polymer consisting essentially of up to 50 head/tail polymerized octapeptides (II) of claim 2.

6. A synthetic polymer consisting essentially of up to 50 head/tail polymerized nonapeptides (III) of claim 2.

7. A synthetic peptide of claim 2 having the sequence

(A) Gly-Dop-Lys

(B) Thr-Gly-Dop-Ser-Ala-Gly-Dop-Lys

(C) Gln-Thr-Gly-Dop-Val-Pro-Gly-Dop-Lys

(D) Gln-Thr-Gly-Dop-Asp-Pro-Gly-Tyr-Lys, or

(E) Gln-Thr-Gly-Dop-Leu-Pro-Gly-Dop-Lys.

8. A synthetic tripeptide according to claim 2, said tripeptide having the sequence

9. A synthetic octapeptide according to claim 2, said octapeptide having the sequence

10. A crosslinked polypeptide which is the reaction product of the components comprising

the synthetic homopolymer or random or block copolymer of claim 3, and

a crosslinking agent.

11. A crosslinked polypeptide according claim 10, wherein said synthetic homopolymer or random or block copolymer is crosslinked through the phenolic groups of said Dop or Tyr or through both said phenolic groups and the pendent --(CH₂ --)₄ NH₂ groups of said Lys.

12. A polypeptide which has been isolated in substantially pure form from the dialyzed DOPA-rich protein or protein precursor stored in the foot of the Atlantic ribbed mussel or which has been isolated from the digested protein obtained from the adhesive protein of the Atlantic ribbed mussel and has been electrophoresed on an acid-urea gel, said polypeptide having the formula:

space="preserve" listing-type="equation">Thr/Ala-Gly-Dop/Tyr-Y.sup.1 -Y.sup.2 -Gly-Dop/Tyr-Lys

or the formula;

space="preserve" listing-type="equation">Gln-Thr/Ala-Gly-Dop/Tyr-Y.sup.1 -Y.sub.2 -Gly-Dop/Tyr-Lys

wherein the above three-letter symbols are defined as follows;

Gln=glutamine residue

Thr=threonine residue

Ala=alanine residue

Gly=glycine residue

Dop=3,4-dihydroxyphenylalanine residue

Tyr=tyrosine residue

Lys=lysine residue, and

Y¹ and Y² represent the following amino acid residues;

Y¹ =a serine, valine, aspartic acid, or leucine residue, and

Y² =an alanine, proline, hydroxyproline, or leucine residue, and

wherein a diagonal mark indicates alternative residues and wherein either said polypeptide can recur up to 20 times.