Protein Engineering, Vol. 16, No. 1, 65-72,
January 2003
© 2003 Oxford University Press
Use of phage display to probe the evolution of binding specificity and affinity in integrins
1 Department of Biochemistry and Biophysics and 2 Bristol-Myers-Squibb Pharmaceutical Company, Wilmington, DE 19880, USA 3 Hematology–Oncology Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 and
4 To whom correspondence should be addressed. E-mail: wdegrado{at}mail.med.upenn.edu
The specific binding of RGD-containing proteins to integrin is a function of both the conformation of and the local sequence surrounding the RGD motif. To study the effect of these factors on integrin binding affinity and specificity, we obtained RGD-containing ligands specific for different integrins presented on the same protein scaffold. The ß-turn region between two anti-parallel ß-strands on the loop I of tendamistat, an inhibitor of 
-amylase, was extended by two residues and randomized in a phagemid library. This library and two subsequently constructed RGD-containing loop I libraries were biopanned with purified integrins IIbß3, Vß3 and Vß5 individually. The sequence analysis of selected tendamistat variants and characterization by phage ELISA revealed that phage adhesion is mediated exclusively by an RGD motif located at only two out of four possible positions on loop I. Further, sequences flanking the RGD motif were specific for different integrin targets. Interestingly, selected tendamistat variants mimic natural integrin ligands, both in sequence similarity and in integrin binding specificity, indicating that various ligand specificity patterns can be generated by driving towards maximum affinity in the integrin–ligand complexes.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. N. Elayadi, K. N. Samli, L. Prudkin, Y.-H. Liu, A. Bian, X.-J. Xie, I. I. Wistuba, J. A. Roth, M. J. McGuire, and K. C. Brown A Peptide Selected by Biopanning Identifies the Integrin {alpha}v{beta}6 as a Prognostic Biomarker for Nonsmall Cell Lung Cancer Cancer Res., June 15, 2007; 67(12): 5889 - 5895. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Sakamoto, Y. Ito, T. Mori, and K. Sugimura Interaction of Human Lactoferrin with Cell Adhesion Molecules through RGD Motif Elucidated by Lactoferrin-binding Epitopes J. Biol. Chem., August 25, 2006; 281(34): 24472 - 24478. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. J. Hosse, A. Rothe, and B. E. Power A new generation of protein display scaffolds for molecular recognition Protein Sci., January 1, 2006; 15(1): 14 - 27. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Holig, M. Bach, T. Volkel, T. Nahde, S. Hoffmann, R. Muller, and R. E. Kontermann Novel RGD lipopeptides for the targeting of liposomes to integrin-expressing endothelial and melanoma cells Protein Eng. Des. Sel., May 1, 2004; 17(5): 433 - 441. [Abstract] [Full Text] [PDF]
|
|