CD36 associates with CD9 and integrins on human blood platelets.
The membrane glycoprotein CD36 is involved in platelet aggregation, inhibition of angiogenesis, atherosclerosis, and sequestration of malaria-parasitized erythrocytes. In this study, immunoprecipitations with anti-CD36 antibodies were performed to identify proteins that associate with CD36 in the platelet membrane. Platelets were solubilized in 1% Triton X-100, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), Brij 96, or Brij 99, and the proteins that coprecipitated with CD36 were identified by peptide mass spectrometry and Western blotting. The tetraspanin protein CD9 and the integrins alphaII(b)beta3 and alpha6beta1 specifically coprecipitated with CD36 from platelets that were solubilized in CHAPS and Brij 99 but not from platelets that were solubilized in Triton X-100. Only CD9 is coprecipitated with CD36 from platelets that were solubilized in Brij 96. Reciprocal immunoprecipitations with antibodies to CD9, alpha6, alphaIIb, or beta3 from Brij 99-solubilized platelets coprecipitated CD36. Coprecipitation of CD36, CD9, and alpha6beta1 was also observed on platelets from a patient with Glanzmann thrombasthenia, indicating that alphaII(b)beta3 is not required for the other proteins to associate. Colocalization of alpha6 and CD36, of CD9 and CD36, and of alpha6 and CD9 was observed on intact platelets prior to solubilization, using double immunofluorescence microscopy. These data indicate that CD36 associates with CD9 and integrins on human blood platelets. These associated proteins may mediate or participate in some of the diverse biological functions of CD36.
Department of Pathology, Beth Israel Deaconess Medical Center, and Harvard Medical School, Boston, MA 02215, USA.
W M
WM
Miao
E
E
Vasile
W S
WS
Lane
J
J
Lawler
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D001792:Blood Platelets / Q000276:immunology / Q000378:metabolism*; D018955:CD36 Antigens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D002793:Cholic Acids / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D006801:Humans; D007118:Immunoassay; D039121:Integrin alpha6beta1; D016023:Integrins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011485:Protein Binding; D020539:Sequence Analysis, Protein; D012995:Solubility / Q000187:drug effects; D060245:Tetraspanin 29
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018955:CD36 Antigens; C554869:CD9 protein, human; D002793:Cholic Acids; D003902:Detergents; D039121:Integrin alpha6beta1; D016023:Integrins; D008562:Membrane Glycoproteins; D060245:Tetraspanin 29; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate