Analysis of the function of activin betaC subunit using recombinant protein.
Activins, TGF-beta superfamily members, have multiple functions in a variety of cells and tissues. Additional activin beta subunit genes, betaC and betaE, have been identified in humans and rodents. To explore the role of activin betaC subunit, we generated recombinant human activin C using Chinese hamster ovary cells. Recombinant activin C from the conditioned medium was purified by consecutive hydrophobic, size-exclusion, and high performance liquid chromatography. SDS-PAGE and Western blot analysis of the purified protein revealed that activin C formed disulfide bridges. However, activin C had no effect on the proliferation of cultured liver cells. Furthermore, there were no significant differences in erythroid differentiation and follicle stimulating hormone secretion in vitro. It was also shown that immunoreactive bands indicated the hetrodimer of activin betaC, and inhibin alpha subunits were detected in the conditioned medium from the activin C-producing cells, which were stably transfected with inhibin alpha subunit cDNA. This suggests that activin betaC subunit may have been present and that it may exert its effect as inhibin C.
Ushiro
Yuuki
Y
Laboratory of Experimental Animal Science, Faculty of Veterinary Medicine, Kitasato University School of Veterinary Medicine and Animal Sciences, Japan.
Hashimoto
Osamu
O
Seki
Machi
M
Hachiya
Akitsu
A
Shoji
Hiroki
H
Hasegawa
Yoshihisa
Y
D000818:Animals; D016466:CHO Cells; D002454:Cell Differentiation; D006224:Cricetinae; D003412:Cricetulus; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D028322:Inhibin-beta Subunits / Q000032:analysis* / Q000502:physiology*; D007265:Inhibins / Q000737:chemistry; D020014:K562 Cells; D008099:Liver / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis* / Q000737:chemistry
C504186:INHBC protein, human; D011994:Recombinant Proteins; C094377:inhibin-alpha subunit; D007265:Inhibins; D005640:Follicle Stimulating Hormone; D028322:Inhibin-beta Subunits