Gonadotropin bioactivities in mouse, hamster, rat and guinea pig pituitaries are largely adsorbed on concanavalin A-sepharose.
The pituitaries of mice, hamsters, guinea pigs and rats were extracted with Tris-Cl buffer and the extracts were chromatographed on Concanavalin A (ConA)-Sepharose into unadsorbed ConA I and adsorbed ConA II fractions. The ConA I fraction was subjected to gel filtration on Sephadex G-100 and fractionated into an unretarded peak and several retarded peaks. The peak with a molecular weight of approximately 40,000 (designated ConAI Sephadex fraction II) was then subjected to ion exchange chromatography on CM-cellulose and fractionated into an unadsorbed CM I and an adsorbed CM II fraction. The ConA II fraction was fractionated by ion exchange chromatography on CM-cellulose into CM I and CM II fractions. The ConA II CM II fraction was the chromatographic fraction which exhibited the highest potency in stimulating testosterone production by isolated rat Leydig cells. Its activity was much higher than the corresponding ConA I Sephadex fraction II CM II fraction which differed chromatographically only by non-adsorption on ConA-Sepharose. The ConA II CM II fraction manifested cross reactivity in a rat luteinizing hormone (LH) radioimmunoassay. The guinea pig pituitary ConA II CM II fraction also cross-reacted in a rat thyroid stimulating hormone radioimmunoassay. The ConA II fractions of hamster and guinea pig pituitary extracts demonstrated follicle stimulating hormone (FSH) activity while the corresponding ConA I fractions did not. The results suggest that the ConA II/ConA II CM II fraction contained most of the FSH and LH activities present in the pituitary extract.
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Department of Biochemistry, Faculty of Medicine, Chinese University of Hong Kong, Shatin, N.T.
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D000818:Animals; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D005260:Female; D005640:Follicle Stimulating Hormone / Q000032:analysis; D006062:Gonadotropins / Q000032:analysis*; D006168:Guinea Pigs; D007985:Leydig Cells / Q000737:chemistry; D007986:Luteinizing Hormone / Q000032:analysis; D008297:Male; D051379:Mice; D010902:Pituitary Gland / Q000737:chemistry*; D011863:Radioimmunoassay; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives / Q000737:chemistry*; D013739:Testosterone / Q000032:analysis; D013972:Thyrotropin / Q000032:analysis
D006062:Gonadotropins; C028326:concanavalin A-sepharose; D013739:Testosterone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D013972:Thyrotropin; D012685:Sepharose