The structure of the beta-propeller domain and C-terminal region of the integrin alphaM subunit. Dependence on beta subunit association and prediction of domains.
The alphaM subunit of integrin Mac-1 contains several distinct regions in its extracellular segment. The N-terminal region has been predicted to fold into a beta-propeller domain composed of seven beta-sheets each about 60 amino acid residues long, with the I-domain inserted between beta-sheets 2 and 3. The structure of the C-terminal region is unknown. We have used monoclonal antibodies (mAbs) as probes to study the dependence of the structure of different regions of the alphaM subunit on association with the beta2 subunit in the alphaM/beta2 heterodimer. All of the mAbs to the I-domain immunoprecipitated the unassociated alphaM precursor and reacted with the alphaM subunit expressed alone on the surface of COS cells. By contrast, four mAbs to the beta-propeller domain did not react with the unassociated alphaM precursor nor with the uncomplexed alphaM subunit expressed on COS cell surface. The four mAbs were mapped to three subregions in three different beta-sheets, making it unlikely that each recognized an interface between the alpha and beta subunits. These results suggest that folding of different beta-propeller subregions is coordinate and is dependent on association with the beta2 subunit. The segment C-terminal to the beta-propeller domain, residues 599-1092, was studied with nine mAbs. A subset of four mAbs that reacted with the alphaM/beta2 complex but not with the unassociated alphaM subunit were mapped to one subregion, residues 718-759, and five other mAbs that recognized both the unassociated and the complexed alphaM subunit were localized to three other subregions, residues 599-679, 820-882, and 943-1047. This suggests that much of the region C-terminal to the beta-propeller domain folds independently of association with the beta2 subunit. Our data provide new insights into how different domains in the integrin alpha and beta subunits may interact.
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Center for Blood Research and Harvard Medical School, Department of Pathology, Boston, Massachusetts 02115, USA.
Oxvig
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Springer
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D019556:COS Cells; D019281:Dimerization; D018604:Epitope Mapping; D005434:Flow Cytometry; D006801:Humans; D016023:Integrins / Q000737:chemistry*; D016177:Macrophage-1 Antigen / Q000737:chemistry*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014162:Transfection / Q000235:genetics
D000911:Antibodies, Monoclonal; D016023:Integrins; D016177:Macrophage-1 Antigen; D011993:Recombinant Fusion Proteins