Study on the partial purification of gonadotropins and thyrotropin from deer (Cervus elaphus) pituitaries

Deer pituitaries were extracted with Tris-Cl buffer and the extract was fractionated on Concanavalin A-Sepharose into an unadsorbed ConA I fraction and an adsorbed ConA II fraction. The ConA II fraction was chromatographed on CM-cellulose to yield an unadsorbed CM I fraction and an adsorbed CM II fraction. The CM I and CM II fractions were then respectively subjected to ion exchange chromatography on DEAE-cellulose to give the resultant unadsorbed DEAE I and adsorbed DEAE II fractions. The ConA II CM II DEAE I fraction was the chromatographic fraction with the highest potency in stimulating testosterone production by isolated rat Leydig cells and the highest immunoreactivity in a rat luteinizing hormone (LH) radioimmunoassay (RIA). The ConA II CM II DEAE II fraction represented the fraction enriched in thyroid stimulating hormone (TSH). It exhibited cross reactivity in a rat TSH RIA and much lower LH bioactivity than the LH-enriched ConA II CM II DEAE I fraction. The ConA II CM I DEAE II fraction represented the follicle stimulating hormone (FSH)-enriched fraction. The ConA I fraction demonstrated negligible FSH immunoreactivity and LH bioactivity, indicating that most of deer FSH and LH were adsorbed on ConA-Sepharose.