Integrin alpha IIb beta 3 (platelet GPIIb-IIIa) recognizes multiple sites in fibronectin.
The binding of fibronectin (Fn) to several integrins involves the Arg-Gly-Asp (RGD) tripeptide sequence. However, linear synthetic RGD peptides do not completely mimic the cell attachment activity of intact Fn or certain large Fn fragments. This suggests that the integrin-Fn interaction involves a more extended surface of Fn than that provided by the RGD sequence. To test this possibility, three novel monoclonal anti-Fn antibodies that inhibit its binding to a purified integrin, alpha IIb beta 3, were developed. The epitopes of these three antibodies mapped to a region at least 55 residues amino-terminal of the RGD sequence. Further, recombinant fragments of Fn containing these epitopes and lacking the RGD site also inhibited the binding of Fn to purified alpha IIb beta 3. These fragments, which spanned Fn residues 1359-1436, bound to alpha IIb beta 3 in a divalent cation-dependent manner. In addition, this region of Fn bound specifically to alpha IIb beta 3 on thrombin-stimulated but not resting platelets. These results demonstrate the presence of additional sequences in Fn that interact with integrin alpha IIb beta 3 and suggest that multiple sites in Fn are involved in its recognition by this integrin.
Bowditch
R D
RD
Committee on Vascular Biology, Scripps Research Institute, La Jolla, California 92037.
Halloran
C E
CE
Aota
S
S
Obara
M
M
Plow
E F
EF
Yamada
K M
KM
Ginsberg
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D004247:DNA; D000939:Epitopes; D005353:Fibronectins / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D015539:Platelet Activation; D011993:Recombinant Fusion Proteins
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D000939:Epitopes; D005353:Fibronectins; D016023:Integrins; D058619:Maltose-Binding Proteins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C047981:arginyl-glycyl-aspartic acid; D004247:DNA