Isolation of porcine follicular fluid inhibin of 32K daltons |
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| Authors: | K Miyamoto Y Hasegawa M Fukuda M Nomura M Igarashi K Kangawa H Matsuo |
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| Affiliation: | 1. Department of Obstetrics & Gynecology, Gunma University School of Medicine, Maebashi, Gunma 371, Japan;2. Department of Biochemistry, Miyazaki Medical College, Kiyotake, Miyazaki 889-16, Japan;1. Departamento de Ecología y Recursos Naturales, Facultad de Ciencias, Universidad Nacional Autónoma de México, Ciudad Universitaria, C.P. 04510 Ciudad de México, Mexico;2. Laboratorio de Herpetología, Departamento de Biología Evolutiva, Facultad de Ciencias, Universidad Nacional Autónoma de México, Ciudad Universitaria, C.P. 04510 Ciudad de México, Mexico;1. Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan;2. Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan;1. Núcleo de Pesquisa e Conservação de Cervídeos (NUPECCE), Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil;2. Laboratório de Estatística Aplicada à Genética e Melhoramento Animal, Departamento de Ciências Exatas, Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil;3. Departamento de Ciências Exatas—Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil;5. Departamento de Zootecnia, Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil;6. Instituto Latino-Americano de Ciências da Vida e da Natureza, Universidade Federal da Integração Latino-Americana (UNILA), Foz do Iguaçu, PR, Brazil;7. Laboratório de Biodiversidade, Conservação e Ecologia de Animais Silvestres (LABCEAS), Universidade Federal do Paraná (UFPR), Curitiba, PR, Brazil;8. Facultad de Medicina Veterinaria y Zootecnia, Universidad Agraria del Ecuador, Guayaquil, Ecuador;1. Department of Orthopedic Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan;2. Department of Orthopedic Biomaterial Science, Graduate School of Medicine, Osaka University, 2-2 Yamadaok, Suita, Osaka 565–0871, Japan |
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| Abstract: | Purification of ovarian inhibin from porcine follicular fluid was performed by using an bioassay based upon the suppression of spontaneous FSH release from cultured cells of rat anterior pituitary. The presence in the follicular fluid of four molecular forms of inhibin activity corresponding to Mr 100K, 80K, 55K and 32K was revealed by SDS-gel electrophoresis under non-reducing conditions. The smallest inhibin amongst them, named 32K inhibin, eliciting about 70% of the total activity in the follicular fluid, was separated by gel filtration in the presence of 8 M urea. By subsequent ion-exchange chromatography, followed by RP-HPLC, 32K inhibin was purified to homogeneity with a 8,000 fold purification factor in a yield of 12%. The purified 32K inhibin was found to comprise two polypeptide subunits (Mr 20K and 13K), linked by disulfide bridges and to specifically suppress the secretion of FSH, but not of LH from the pituitary cells. |
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