Topographic analysis of human follicle-stimulating hormone-beta using anti-peptide antisera.
The purpose of this study was to identify peptide sequences of human follicle-stimulating hormone-beta (hFSH beta) which are accessible subsequent to association with hFSH alpha in heterodimeric hFSH. Antisera were raised against synthetic peptides (Abpep) corresponding to hFSH beta sequences 1-20, 16-36, 33-53, 49-67, 66-85, 81-100 and 98-111. The topography of hFSH beta was studied by testing the binding of these antisera to hFSH beta and hFSH captured by monoclonal antibodies (MAb) in an enzyme-linked immunosorbent assay (ELISA). When hFSH and hFSH beta were captured by the same MAb, binding of Ab16-36, Ab33-53, Ab81-100 and Ab98-111 to hFSH was significantly lower compared to hFSH beta. However, compared to other Abpep, binding of Ab35-53 to hFSH was strong. Similar results were obtained when hFSH was captured by an alpha-specific MAb (10.3A6). Using 10.3A6, it was also possible to demonstrate significant binding of Ab49-67 to hFSH. The data suggests that residues in regions 33-53 and 49-67 of hFSH beta appear to be accessible in the heterodimeric hFSH in addition to the glycosylated region of 1-15. Regions 16-36, 33-53, 81-100 and 98-111 of hFSH beta appear to contain subunit contact-associated sequences which are either masked or structurally altered subsequent to association with hFSH alpha in the heterodimeric hFSH.
Vakharia
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DD
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Bryant
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SH
Dias
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JA
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000276:immunology; D037201:Follicle Stimulating Hormone, beta Subunit; D015292:Glycoprotein Hormones, alpha Subunit / Q000737:chemistry / Q000276:immunology; D006801:Humans; D010455:Peptides / Q000276:immunology; D011817:Rabbits
D000906:Antibodies; D000911:Antibodies, Monoclonal; D037201:Follicle Stimulating Hormone, beta Subunit; D015292:Glycoprotein Hormones, alpha Subunit; D010455:Peptides; D005640:Follicle Stimulating Hormone