Identification of palmitoylation sites on CD4, the human immunodeficiency virus receptor.
We report that the cell surface glycoprotein CD4 expressed in HeLa cells can be metabolically labeled with [3H]palmitic acid. Analysis of the 3H-label after hydrolysis of the protein indicated that it was incorporated predominantly as palmitic acid. Comparison of the amount of [3H]palmitate incorporated into CD4 with that incorporated into a protein known to contain one molecule of esterified palmitate suggested that one to two molecules of palmitate were added to CD4. The fatty acid was readily cleaved from CD4 by treatment with weak base suggesting a thioester linkage. Mutations of each of 2 cysteine residues, Cys394 and Cys397, in CD4 at the junction of the transmembrane and cytoplasmic domains reduced labeling with [3H]palmitic acid, and mutation of both cysteines eliminated labeling. These results indicate that both cysteines are esterified to palmitate. Modification with palmitate was not required for expression of CD4 on the cell surface or for binding of p56lck to its cytoplasmic domain.
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Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510.
Rose
J K
JK
D000595:Amino Acid Sequence; D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics / Q000378:metabolism*; D002853:Chromatography, Liquid; D003545:Cysteine / Q000378:metabolism; D006678:HIV / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism*; D010957:Plasmids; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational
D015704:CD4 Antigens; D010169:Palmitic Acids; D019308:Palmitic Acid; D008715:Methionine; D003545:Cysteine