Endocrinology, Vol 124, 923-929, Copyright © 1989 by Endocrine Society

ARTICLES

Monoclonal antibodies directed to human and equine chorionic gonadotropins as probes for the topographic analysis of epitopes on the human alpha-subunit

JM Bidart, F Troalen, GR Bousfield, C Bohuon and D Bellet
Unite de Biochimie Clinique, Institut Gustave-Roussy, Villejuif, France.

To improve our knowledge of the structural features of the alpha- subunit of hCG we have studied the antigenic site recognized by monoclonal antibody (MAb) ECG01 raised against equine CG (eCG) which binds to hormones and alpha-subunits from human and equine species. We have also delineated regions of hCG alpha comprising the epitope recognized by HT13 which was raised against hCG and binds to hCG and hCG alpha. To define the residues involved in the antigenic sites recognized by ECG01 and HT13, we have studied the reactivities of these two MAbs with native or chemically modified LH and CG with subunits from equine, human, or ovine (o) species or with synthetic peptides analogous to various portions of hCG alpha. We have also compared these reactivities with those displayed by MAbs AHT20 and FA 36, whose epitopes have been previously described; anti-hCG alpha MAb AHT20 is specific for the free alpha-subunits of various species and recognizes residues localized to the 36-41 region of hCG alpha, whereas antipeptide MAb FA36 binds to the 87-92 carboxyl-terminal part of hCG alpha. Our results show that the epitopes of HT13 and ECG01 are 1) probably discontinuous, as these MAbs did not bind to the reduced and S- carboxymethylated hCG alpha; and 2) constituted by residues borne on the 1-35 and 52-86 sequences, as they do recognize the hCG alpha core missing the 36-51 portion, yet do not recognize hCG alpha-(87-92) region recognized by FA36. The comparative studies performed with specific two-site immunoradiometric assays to determine the interspecies cross-reactivities of the MAbs allow us hypothetical assignment of residues on the primary structure of hCG alpha. The antigenic site recognized by ECG01 might include two to six amino acids, four of these residues being located at inverted places compared to those of oLH alpha (Asp6/Gly22 and Arg67/Lys75). These residues present important charged functional groups highly conserved among evolutionarily related variants, and it is likely that they are located on the surface of both the intact hormone and its alpha-subunit. Three peptidic portions of hCG alpha, 16-17, 64-66, and 73-76, respectively, might be involved in the epitope recognized by HT13, although we could not rule out the possibility that other residues were also involved in the antigenic site. These observations allow us to identify several residues as potentially constituting the epitopes recognized by two MAbs on both hCG and hCG alpha.

This article has been cited by other articles:

				W. J. Walton, V. T. Nguyen, V. Y. Butnev, V. Singh, W. T. Moore, and G. R. Bousfield Characterization of Human FSH Isoforms Reveals a Nonglycosylated {beta}-Subunit in Addition to the Conventional Glycosylated {beta}-Subunit J. Clin. Endocrinol. Metab., August 1, 2001; 86(8): 3675 - 3685. [Abstract] [Full Text] [PDF]

				L. Cosowsky, S. N. V. Rao, G. J. Macdonald, H. Papkoff, R. K. Campbell, and W. R. Moyle The Groove between the alpha- and beta-Subunits of Hormones with Lutropin (LH) Activity Appears to Contact the LH Receptor, and Its Conformation Is Changed during Hormone Binding J. Biol. Chem., August 25, 1995; 270(34): 20011 - 20019. [Abstract] [Full Text] [PDF]

				W. R. Moyle, R. K. Campbell, S. N. V. Rao, N. G. Ayad, M. P. Bernard, Y. Han, and Y. Wang Model of Human Chorionic Gonadotropin and Lutropin Receptor Interaction That Explains Signal Transduction of the Glycoprotein Hormones J. Biol. Chem., August 25, 1995; 270(34): 20020 - 20031. [Abstract] [Full Text] [PDF]