title: Immunoelectron microscopy studies with a monoclonal antibody directed against a receptor recognition site epitope in human alpha 2-macroglobulin.
abstract: Alpha 2-Macroglobulin (alpha 2M) is a plasma proteinase inhibitor that binds up to 2 mole of proteinase per mole of inhibitor. Proteinase binding or reaction with small primary amines causes a major conformational change in alpha 2M. As a result of this conformational change, a new epitope recognized by monoclonal antibody 7H11D6 is exposed. The association of alpha 2M-proteinase or alpha 2M-methylamine with alpha 2M cellular receptors is prevented by 7H11D6. In this investigation, the binding of 7H11D6 to alpha 2M was studied by electron microscopy. 7H11D6 bound to alpha 2M-methylamine and alpha 2M-trypsin but not to native alpha 2M. The structure of alpha 2M after conformational change resembled the letter "H." 7H11D6 epitopes were identified near the apices of the four arms in the alpha 2M "H" structure. 7H11D6 that was adducted to colloidal gold (7HAu) retained the specificity of the free antibody (binding to alpha 2M-trypsin but not to native alpha 2M). alpha 2M conformational change intermediates prepared by sequential reaction with a protein crosslinker and trypsin also bound 7HAu. These results suggest that a complete alpha 2M conformational change is not necessary for 7H11D6 epitope exposure and may not be required for receptor recognition. 7HAu was used to isolate a preparation consisting primarily of binary alpha 2M-trypsin (1 mole trypsin per mole alpha 2M instead of 2). Structures resembling the letter "H" were most common; however, each field showed some atypical molecules with arms that were compacted instead of thin and elongated.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Pathology, University of Virginia Health Sciences Center, Charlottesville 22908.
authors.forename: N L
authors.initials: NL
authors.lastname: Figler
authors.forename: D K
authors.initials: DK
authors.lastname: Strickland
authors.forename: M
authors.initials: M
authors.lastname: Allietta
authors.forename: S L
authors.initials: SL
authors.lastname: Gonias
mesh_terms: D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D002945:Cisplatin; D000939:Epitopes; D006801:Humans; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D000511:alpha-Macroglobulins / Q000276:immunology / Q000648:ultrastructure*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D000511:alpha-Macroglobulins; D002945:Cisplatin