Folding of the conserved domain but not of flanking regions in the integrin beta2 subunit requires association with the alpha subunit.
We have used immunoprecipitation with mAbs to probe folding during biosynthesis of the beta2 integrin subunit of lymphocyte function-associated antigen 1 (LFA-1; CD11a/CD18) before and after association with the alphaL subunit. An evolutionarily conserved region is present in the beta2 subunit between amino acid residues 102 and 344. mAbs to one subregion before the conserved region, and two subregions after the conserved domain, immunoprecipitated both the unassociated beta'2 precursor and mature alphaL/beta2 complex, suggesting portions of these subregions are folded before association with alphaL. An activating mAb to the C-terminal cysteine-rich region, KIM127, preferentially bound to the unassociated beta subunit, suggesting that it may bind to an epitope that is in an alphabeta interface in unactivated LFA-1. By contrast, mAbs to five different epitopes in the conserved region did not react with unassociated beta'2 precursor, suggesting that this region folds after alphaL association and is intimately associated with the alphaL subunit in the alphaL/beta2 complex. mAbs to two different epitopes that involve the border between the conserved region and the C-terminal segment, were fully or partially reactive with the beta'2 precursor, suggesting that this region is partially folded before association with alphaL. The findings suggest that the conserved region is a distinct folding and hence structural unit, and is intimately associated with the alpha subunit.
Huang
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Department of Pathology, Harvard Medical School, Boston, MA 02115, USA.
Lu
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Springer
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D018821:CD18 Antigens / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D017124:Conserved Sequence; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D017510:Protein Folding
D000911:Antibodies, Monoclonal; D018821:CD18 Antigens