Our previous studies showed that the alpha 5 beta 1 integrin selects cysteine pair-containing RGD peptides from a phage display library based on a random hexapeptide. We have therefore searched for more selective peptides for this integrin using a larger phage display library, where heptapeptides are flanked by cysteine residues, thus making the inserts potentially cyclic. Most of the phage sequences that bound to alpha 5 beta 1 (69 of 125) contained the RGD motif. Some of the heptapeptides contained an NGR motif. As the NGR sequence occurs in the cell-binding region of the fibronectin molecule, this sequence could contribute to the specific recognition of fibronectin by alpha 5 beta 1. Selection for high affinity peptides for alpha 5 beta 1 surprisingly yielded a sequence RRETAWA that does not bear obvious resemblance to known integrin ligand sequences. The synthetic cyclic peptide GACRRETAWACGA (*CRRETAWAC*) was a potent inhibitor of alpha 5 beta 1-mediated cell attachment to fibronectin. This peptide is nearly specific for the alpha 5 beta 1 integrin, because much higher concentrations were needed to inhibit the alpha v beta 1 integrin, and there was no effect on alpha v beta 3- and alpha v beta 5-mediated cell attachment to vitronectin. The peptide also did not bind to the alpha IIb beta 3 integrin. *CRRETAWAC* appears to interact with the same or an overlapping binding site in alpha 5 beta 1 as RGD, because cell attachment to *CRRETAWAC* coated on plastic was divalent cation dependent and could be blocked by an RGD-containing peptide. These results reveal a novel binding specificity in the alpha 5 beta 1 integrin.
Skip Nav Destination
Article navigation
1 February 1994
Article|
February 01 1994
Isolation of a highly specific ligand for the alpha 5 beta 1 integrin from a phage display library
E Koivunen,
E Koivunen
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Search for other works by this author on:
B Wang,
B Wang
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Search for other works by this author on:
E Ruoslahti
E Ruoslahti
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Search for other works by this author on:
E Koivunen
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
B Wang
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
E Ruoslahti
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1994) 124 (3): 373–380.
Citation
E Koivunen, B Wang, E Ruoslahti; Isolation of a highly specific ligand for the alpha 5 beta 1 integrin from a phage display library. J Cell Biol 1 February 1994; 124 (3): 373–380. doi: https://doi.org/10.1083/jcb.124.3.373
Download citation file:
Sign in
Don't already have an account? Register
Client Account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Sign in via your Institution
Sign in via your InstitutionSuggested Content
Email alerts
Advertisement
Advertisement