Identification of a region in the integrin beta3 subunit that confers ligand binding specificity.
Many integrin adhesion receptors bind ligands containing the Arg-Gly-Asp (RGD) peptide motif. Most integrins exhibit considerable specificity for particular ligands and can distinguish among the many conformations of RGD. In this study we identify the domain of the integrin beta subunit involved in determining ligand binding specificity. Chimeras of beta3 and beta5, the most homologous integrin beta subunits, were expressed with alphav on the surface of human 293 cells. The ligand binding phenotype of each chimera was assessed using the ligands Fab-9 and fibrinogen, both of which have a binding preference for alphavbeta3. The results of the study show that when exons C and D of the beta3 subunit (residues 95-233) are substituted into beta5, the chimera gained the ability to bind Fab-9 with an affinity close to that of wild-type alphavbeta3. This chimera was able to mediate cell adhesion to fibrinogen. Furthermore, the swap of only a 39-residue segment of this larger domain, beta3 residues 164-202, into the backbone of beta5 enabled the chimeric integrin to bind soluble Fab-9. This small domain is highly divergent among the integrin beta subunits, suggesting that it may play a role in determining ligand selection by all integrins.
Lin
E C
EC
Program on Cell Adhesion and the Extracellular Matrix, The Burnham Institute, La Jolla Cancer Research Center, La Jolla, California 92037, USA.
Ratnikov
B I
BI
Tsai
P M
PM
Carron
C P
CP
Myers
D M
DM
Barbas
C F
CF
Smith
J W
JW
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D002460:Cell Line; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D039661:Integrin beta3; D008024:Ligands; D008969:Molecular Sequence Data; D010980:Platelet Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D019096:Vitronectin / Q000378:metabolism
D015703:Antigens, CD; D039661:Integrin beta3; D008024:Ligands; D010980:Platelet Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D019096:Vitronectin; D005340:Fibrinogen