title: Differential thermal stability of human, bovine and ovine Follicle-Stimulating Hormone (FSH) and Luteinizing Hormone (LH) quaternary structures.
abstract: Quaternary structure of human, bovine and ovine Follicle-Stimulating Hormones (hFSH, bFSH and oFSH) and Luteinizing Hormone was assessed in sandwich ELISAs using monoclonal anti-oFSHβ or anti-oLHβ antibodies, respectively, for capture and a biotinylated anti-hFSHα (α4 epitope) for detection. Neither free subunit gave any signal in this assay so that it was possible to measure the residual heterodimeric fraction after thermal treatment of the gonadotropins under study. The hormones were subjected to 5-min heating between 37 and 90 °C before rapid cooling in melting ice before ELISA. The data show half-dissociation of natural and recombinant human and ovine FSH preparations between 68 and 74 °C whereas bovine FSH preparations exhibited lower stability in these conditions with half-dissociation between 61 and 64 °C. Moreover, whereas all human and bovine as well as most ovine FSH preparations were fully dissociated at temperatures above 80 °C, one natural oFSH and one recombinant hLH preparations contained an important fraction that resisted dissociation even at 93 °C and retained in vitro bioactivity. This suggests the existence of gonadotropin αβ heterodimer with covalently linked subunits. Similarly, about 20% of the recombinant hLH preparation was also found withstand heat denaturation and also probably to have cross-linked subunits. The origin and chemical nature of these inter-subunit bonds remain to be determined.
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Maya
authors.initials: M
authors.lastname: Haj Hassan
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Cahoreau
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Gwenhael
authors.initials: G
authors.lastname: Jégot
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Jouanny
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Mariot
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: François
authors.initials: F
authors.lastname: Lecompte
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France.
authors.forename: Danièle
authors.initials: D
authors.lastname: Klett
authors.affiliation: Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS), Unit «Physiologie de la Reproduction et des Comportements», 37380 Nouzilly, France. Electronic address: combarno@tours.inra.fr.
authors.forename: Yves
authors.initials: Y
authors.lastname: Combarnous
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001681:Biological Assay; D002417:Cattle; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000378:metabolism*; D057809:HEK293 Cells; D006358:Hot Temperature; D006801:Humans; D007985:Leydig Cells; D007986:Luteinizing Hormone / Q000737:chemistry* / Q000378:metabolism*; D008297:Male; D051379:Mice; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone
keywords: ELISA; Gonadotropin; Reproduction; Structure–function