The functional cell surface glycoprotein CD9 is distinguished by being the major fatty acid acylated and a major iodinated cell-surface component of the human platelet.
We showed that a 22 kDa protein (which comigrated with the leukocyte differentiation antigen CD9 as determined by immunoblotting with the platelet-activating mAb 50H.19) is a major iodinated component of the platelet surface. The iodinated protein was identified as CD9 by limited proteolysis analysis. The major acylated protein in platelets incubated with [3H]palmitic acid also had a mobility of 22 kDa. The radiolabelled fatty acid in CD9 appears to be ester bonded, as it is removed by treatment with hydroxylamine. Non-enzymatic ligation of the fatty acid is not involved. Since platelets lack protein synthetic capacity, the palmitolation of a surface protein indicates the existence of a plasma-membrane located transacylase which functions independently of protein synthesis. Limited proteolysis analysis of the palmitylated protein obtained by immunoprecipitation with mAb 50H.19 confirmed its identity as CD9. An additional novel minor component of 27 kDa was detected in platelets by immunoprecipitation of 125I-surface-labelled, or [3H]palmitic acid-labelled protein, and by immunoblotting with mAb 50H.19. The analogous cleavage patterns obtained by the limited proteolysis analysis of the 22, 24 and 27 kDa glycoproteins suggest that they may be differently modified variants of a single polypeptide.
Seehafer
J G
JG
Department of Medicine, Cross Cancer Institute, Edmonton, Canada.
Slupsky
J R
JR
Tang
S C
SC
Shaw
A R
AR
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000954:Antigens, Surface / Q000032:analysis*; D001792:Blood Platelets / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis / Q000276:immunology; D006801:Humans; D007962:Leukocytes / Q000276:immunology; D016169:Lymphocyte Function-Associated Antigen-1; D008562:Membrane Glycoproteins; D008970:Molecular Weight; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*; D013601:T-Lymphocytes / Q000276:immunology; D060245:Tetraspanin 29
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000954:Antigens, Surface; C554869:CD9 protein, human; D016169:Lymphocyte Function-Associated Antigen-1; D008562:Membrane Glycoproteins; D010980:Platelet Membrane Glycoproteins; D060245:Tetraspanin 29