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Binding of the Adhesive Protein Complex (LFA‐1/Mac‐1/p150,95) to Concanavalin A
Author(s) -
Schmalstieg Frank C.,
Rudloff Helen E.,
Anderson Donald C.
Publication year - 1986
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.39.2.193
Subject(s) - concanavalin a , biology , glycoprotein , immunoprecipitation , gel electrophoresis , sodium dodecyl sulfate , protein subunit , microbiology and biotechnology , membrane protein , g alpha subunit , biochemistry , electroblotting , membrane glycoproteins , polyacrylamide gel electrophoresis , silver stain , membrane , enzyme , in vitro , gene
At least 30 proteins from human PMNL plasma membranes capable of binding concanavalin A (Con A), can be identified after surface labeling with 125 I and subsequent sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). Immunoprecipitation of the labeled proteins after solubilization in non‐ionic detergents, with monoclonal antibodies (MAbs) directed against a family of leukocyte membrane proteins (LFA‐1, Mac‐1, p150,95, and the beta subunit these glycoproteins share), indicated that Mac‐1 and p150,95 were bound by Con A. Dissociation of the alpha and beta subunits with sodium dodecyl sulfate, electrophoresis, transfer to nitrocellulose paper, and subsequent binding of these proteins by Con A demonstrated Con A retention by Mac‐1‐ α , p150,95‐ α , and the common beta subunit. Affinity of Con A for LFA‐1‐ α from human peripheral blood PMNL could not be confirmed by direct binding or electroblotting. Similar experiments in a patient deficient in LFA‐1, Mac‐1, p150,95, and the beta subunit confirmed that Mac‐1‐ α and the beta subunit were important Con A‐binding proteins.