The mouse leukocyte adhesion proteins Mac-1 and LFA-1: studies on mRNA translation and protein glycosylation with emphasis on Mac-1.
Translation in vitro of mRNA and immunoprecipitation with specific rabbit antisera showed that the unglycosylated precursor polypeptides of the mouse Mac-1 and lymphocyte function associated antigen (LFA-1) alpha subunits are 130,000 Mr and 140,000 Mr, respectively. Furthermore, polysomes purified by using anti-Mac-1 IgG yielded a similar major product of translation in vitro of Mr = 130,000. The Mac-1 and LFA-1 alpha subunit translation products are immunologically noncross-reactive, showing that differences between these related proteins are not due to post-translational processing. Mac-1 and LFA-1 alpha subunits could only be in vitro translated from mRNA from cell lines the surfaces of which express the corresponding Mac-1 and LFA-1 alpha-beta complexes, showing tissue-specific expression is regulated at the mRNA level. The glycosylation of Mac-1 was examined by both translation in vitro in the presence of dog pancreas microsomes and by biosynthesis in vivo and treatment with tunicamycin, endoglycosidase H, and the deglycosylating agent trifluoromethane sulfonic acid. High mannose oligosaccharides are added to the Mac-1 alpha and beta polypeptide backbones of Mr = 130,000 and 72,000, respectively, to yield precursors of Mr = 164,000 and 91,000, respectively. The alpha and beta subunit precursors are then processed with partial conversion of high mannose to complex type carbohydrate to yield the mature subunits of Mr = 170,000 and 95,000, respectively.
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis* / Q000096:biosynthesis* / Q000302:isolation & purification; D002448:Cell Adhesion; D004285:Dogs; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism*
D000954:Antigens, Surface; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D009844:Oligosaccharides; D011498:Protein Precursors; D012333:RNA, Messenger