Modified integrin polypeptides, modified integrin polypeptide dimers, and uses thereof
AssigneeChildren's Medical Center Corporation
InventorsTimothy Alan Springer, Xianchi Dong, Chafen Lu
Life of this patent
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Abstract
Described herein are modified integrin α and/or β headpiece polypeptides, and crystallizable integrin polypeptide dimers comprising a modified integrin α and/or β headpiece polypeptide and a disulfide bond linking the two integrin headpiece polypeptide subunits. Methods for using the modified integrin α and/or β headpiece polypeptides and the integrin polypeptide dimers are also provided herein. For example, methods for characterizing integrin-ligand interaction and identifying integrin ligands are also provided herein. In some embodiments, the identified integrin ligands can be used as inhibitors of integrins.
Claims16
A modified integrin αv headpiece polypeptide comprising an amino acid sequence of SEQ ID NO:
- 1 with at least one Cys residue introduced thereto by one or more of the following modifications (a)-(e):
- a. substitution of amino acid residues 399-401 with one of the following:
- (i) Ser-Cys-Pro;
- (ii) Gly-Cys-Pro;
- (iii) Ser-Cys-Gly;
- (iv) Gly-Cys-Gly;
- (v) Ser-Gly-Cys-Pro (SEQ ID NO:
- 59);
- (vi) Ser-Cys-Gly-Pro (SEQ ID NO:
- 60);
- (vii) Gly-Cys-Gly-Pro (SEQ ID NO:
- 61);
- and (viii) Ser-Gly-Cys-Gly (SEQ ID NO:
- 62);
- b. substitution of amino acid residues 310-311 (Gln-Glu) with Gly-Cys;
- c. substitution of amino acid residues 299 (Leu) and 310 (Gln) with Cys and Gly, respectively;
- d. substitution of amino acid residues 302-311 (Asp-Arg-Gly-Ser-Asp-Gly-Lys-Leu-Gln-Glu) (SEQ ID NO:
- 63) with Gly-Gln-Gly-Cys (SEQ ID NO:
- 64);
- and/or
- e. substitution of amino acid residue 299 (Leu) to Cys and substitution of amino acid residues 302-310 (Asp-Arg-Gly-Ser-Asp-Gly-Lys-Leu-Gln) (SEQ ID NO:
- 65) with Gly-Gln-Gly.
A modified integrin β6 headpiece polypeptide comprising a βI domain of integrin β6 subunit with at least one Cys residue introduced thereto by one or more of the following modifications (a)-(b), the βI domain is defined from residues DYP to residues ELR as shown in an amino acid sequence of SEQ ID NO:
- 2:
- a. substitution of amino acid residue 294 (Thr) with Cys;
- and
- b. substitution of amino acid residue 296 (Gly) with Cys.
A modified integrin β3 headpiece polypeptide comprising amino acid residues 27 to 498 of SEQ ID NO:
- 5 with at least one Cys residue introduced thereto by substitution of amino acid residue 293 (Gln) with Cys.
A modified integrin β8 headpiece polypeptide comprising amino acid residues 43 to 498 of SEQ ID NO:
- 6 with at least one Cys residue introduced thereto by substitution of amino acid residue 301 (Val) with Cys.
A modified integrin αv headpiece polypeptide comprising an amino acid sequence of SEQ ID NO:
- 131.
A modified integrin β6 headpiece polypeptide comprising an amino acid sequence of SEQ ID NO:
- 133 (with or without a His6 tag (SEQ ID NO:
- 132)) or SEQ ID NO:
- 135.
A modified integrin β3 headpiece polypeptide comprising an amino acid sequence of SEQ ID NO:
- 136.
A modified integrin β8 headpiece polypeptide comprising an amino acid sequence of SEQ ID NO:
- 134 or SEQ ID NO:
- 137.
Dependencies are inferred from the claim text.