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Physicochemical and biological characterizations of pregnant mare serum gonadotropin and its subunits
Affiliation:1. Hormone Research Laboratory, University of California, San Francisco, Calif. 94143 U.S.A.;2. Reproductive Endocrinology Center, University of California, San Francisco, Calif. 94143 U.S.A.;1. Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research “Campus Mare Nostrum”, University of Murcia, 30100, Murcia, Spain;2. Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de La Salud, Carretera Buenavista S/n, 30120 El, Palmar, Murcia, Spain;3. Department of Clinical & Experimental Medicine (IKE), Linköping University, Campus US, 58183, Linköping, Sweden;4. Salk Institute for Biological Studies, 10010 N. Torrey Pines Rd., La Jolla, CA, 92037, USA;1. Institute of Endocrinology, Prague, Czech Republic;2. National Institute of Mental Health, Klecany, Czech Republic;1. Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350108, PR China;2. Cellular and Developmental Engineering Center, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350108, PR China;1. Department of Obstetrics and Gynecology, Ernst-Moritz-Arndt-University, D-17475 Greifswald, Germany;2. Department of Obstetrics & Gynecology, Justus-Liebig-University, D-35385 Giessen, Germany;3. Department of Anesthesiology, Justus-Liebig-University, D-35385 Giessen, Germany;4. Institute for Clinical Immunology & Transfusion Medicine, Justus-Liebig-University, D-35385 Giessen, Germany;5. Department of Biotechnology, School of Life Sciences, Central University of Rajasthan NH-8, Bandar Sindri, Distt. Ajmer, Rajasthan 305801, India;6. Institute of Biochemistry, Medical School, Justus-Liebig-University, D-35392 Giessen, Germany;1. Laboratorio de InmunoFisiología, Grupo Fisiopatología de la mujer, del embarazo, parto y puerperio. Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain;2. Servicio de Cirugía General, Sección Cirugía Endocrino-Metabólica, Hospital General Universitario Gregorio Marañón, Madrid, Spain;3. Animalario, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain;4. Servicio de Ginecología, Hospital General Universitario Gregorio Marañón, Spain;5. Laboratorio de Inmuno-oncología, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain;6. Unidad de Microscopía Confocal, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain;1. ETH Zurich, Animal Physiology, Institute of Agricultural Sciences, Zurich, Switzerland;2. Technische Universität München, Physiology Weihenstephan, Freising, Germany;3. University of Veterinary Medicine Hannover, Institute of Anatomy, Hannover, Germany;4. University of Veterinary Medicine Hannover, Unit for Reproductive Medicine, Hannover, Germany;5. Utrecht University, Dep. of Equine Sciences, Faculty of Vet. Med., Utrecht, The Netherlands
Abstract:
Pregnant mare serum gonadotropin and its subunits have been further characterized. Ultracentrifugation of the gonadotropin at pH 1.3 and 11.5 showed little evidence of dissociation compared to pH 8.2. Highly purified subunits are obtained by urea dissociation and ion-exchange chromatography followed by gel-filtration. Circular dichroism spectra of the gonadotropin and its subunits are much like those of ovine lutropin and its subunits in that there is little evidence for secondary structure and one or more tyrosine residues are inaccessible in the intact gonadotropin compared to the subunits. The α-subunit possesses almost 3 times as much total carbohydrate as the β-subunit; the individual sugar composition of each was determined as well as the amino acid composition. The α-subunit begins with the sequence NH2-Phe-Pro (Gly or Pro) … and terminates with isoleucine. The β-subunit has the sequence NH2-Ser-Pro-Gly …; no C-terminal residue is detectable by either carboxypeptidase or hydrazinolysis. Biological studies show the gonadotropin to be active in assays specific for both lutropin and follitropin. Precipitin test in agar with rabbit antiserum against the gonadotropin show that the beta subunit cross-reacts whereas the alpha subunit does not.
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