A mitogenic fibrinogen receptor that differs from glycoprotein IIb-IIIa. Identification by affinity chromatography and by covalent cross-linking.
The mitogenic effect of human fibrinogen on the hemopoietic cell lines Raji and JM is mediated by a specific receptor with biochemical and functional properties different from those of the platelet fibrinogen receptor, the glycoprotein complex IIb-IIIa (Levesque, J.P., Hatzfeld, A., and Hatzfeld, J. (1986) Proc. Natl. Acad. Sci. U.S. A. 83, 6494-6498; Levesque, J.P., Hatzfeld, A., Hudry-Clergeon, G., Wilner, G.D., and Hatzfeld, J. (1987) J. Cell. Physiol. 232, 303-310). This work describes the identification of the mitogenic fibrinogen receptor (MFR) by two independent methods, affinity chromatography and covalent cross-linking. Affinity chromatography of surface-labeled cell extracts on fibrinogen-Sepharose revealed a 94 +/- 2-kDa membrane protein that bound specifically to fibrinogen-Sepharose only on cells that expressed the MFR. Its molecular mass was not modified after reduction. This was confirmed by cross-linking fibrinogen to surface-labeled Raji cells using the cleavable cross-linkers, ethyleneglycobis(succinimidyl succinate) and dithiobis(succinimidyl propionate). Complexes between fibrinogen and iodinated cell membrane proteins were immunoprecipitated by anti-fibrinogen antibodies. The biochemical cleavage of these immunoprecipitated conjugates gave rise to a 92 +/- 3-kDa membrane protein whose molecular mass was not modified after reduction. We conclude that fibrinogen binds specifically to a 92-94-kDa MFR which does not belong to the integrin family.
Laboratoire de Biologie Cellulaire et Moléculaire, Institut de Cancérologie et d'Immunogénetique (Centre National de la Recherche Scientifique) Hôpital Paul-Brousse, Villejuif, France.
J P
JP
Levesque
J
J
Hatzfeld
A
A
Hatzfeld
D002460:Cell Line; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D013388:Succinimides
D003432:Cross-Linking Reagents; D010980:Platelet Membrane Glycoproteins; D013388:Succinimides; C028722:ethylene glycolylbis(succinimidyl succinate); D005340:Fibrinogen; C011240:dithiobis(succinimidylpropionate)