Monoclonal antibodies to the insulin receptor stimulate the intrinsic tyrosine kinase activity by cross-linking receptor molecules.
The effect of monoclonal anti-insulin receptor antibodies on the intrinsic kinase activity of solubilized receptor was investigated. Antibodies for six distinct epitopes stimulated receptor autophosphorylation and kinase activity towards exogenous substrates. This effect of antibodies was seen only within a narrow concentration range and monovalent antibody fragments were ineffective. Evidence was obtained by sucrose density-gradient centrifugation for the formation of antibody-receptor complexes which involved both inter- and intra-molecular cross-linking, although stimulation of autophosphorylation appeared to be preferentially associated with the latter. There was partial additivity between the effects of insulin and antibodies in stimulating autophosphorylation, although the sites of phosphorylation appeared identical on two-dimensional peptide maps. Antibodies for two further epitopes failed to activate receptor kinase, but inhibited its stimulation by insulin. The effects of antibodies on kinase activity paralleled their metabolic effects on adipocytes, except for one antibody which was potently insulin-like in its metabolic effects, but which antagonized insulin stimulation of kinase activity. It is concluded that antibodies activate the receptor by cross-linking subunits rather than by reacting at specific epitopes. The ability of some antibodies to activate receptor may depend on receptor environment as well as the disposition of epitopes.
O'Brien
R M
RM
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, UK.
Soos
M A
MA
Siddle
K
K
D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000201:enzymology; D003432:Cross-Linking Reagents; D005260:Female; D006801:Humans; D007700:Kinetics; D010449:Peptide Mapping; D010766:Phosphorylation; D010920:Placenta / Q000201:enzymology; D011505:Protein-Tyrosine Kinases / Q000276:immunology / Q000378:metabolism*; D011972:Receptor, Insulin / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin