Antigenic, receptor-binding and mitogenic activity of proteolytic fragments of human growth hormone.
Analysis of the subtilisin-digested, two-chain form of human growth hormone (hGH) and its constituent polypeptide fragments has been aided by the use of monoclonal antibodies which bind specifically to four distinct epitopes on the native hormone. Using the SDS-polyacrylamide immunoblotting technique, it was shown that one epitope (shared with human chorionic somatomammotropin) detected by EB1 (or EB3) antibody was expressed to a similar extent by both the N-terminal (15 K) and C-terminal (7 K) polypeptides. This epitope is unique in that it represents a repeating determinant within the single chain structure of the hormone. Another three epitopes detected by monoclonal antibodies QA68/NA27, NA71 or NA39/EB2 were absent from the 7-K fragment but were expressed on the 15-K fragment to a similar extent to that on unmodified growth hormone. Binding of NA71 antibody was demonstrated only by radioimmunoassay since this, presumably conformational epitope could not be detected by immunoblotting. The functional importance of the 15-K peptide was demonstrated by its ability to bind specifically to hormone receptors on IM9 human lymphoblastoid cells and by its retention of mitogenicity for the NB2 rat lymphoma cell line. However, all tested monoclonal antibodies inhibited the binding of [125I]15-K to IM9 cell receptors by either steric hindrance or by an allosteric mechanism and therefore could not be further related topographically to the receptor-binding moiety of hGH.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002455:Cell Division / Q000187:drug effects; D000939:Epitopes / Q000276:immunology; D013006:Growth Hormone / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008214:Lymphocytes / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology*; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism; D011986:Receptors, Somatotropin; D013381:Subtilisins
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011986:Receptors, Somatotropin; D013006:Growth Hormone; D013381:Subtilisins