Effective separation of human chorionic gonadotropin subunits by high performance gel permeation chromatography.
Human chorionic gonadotropin (hCG) was effectively separated into alpha and beta subunits by treating with 8 M guanidine hydrochloride, followed by high performance gel permeation chromatography (GPC) on a 2.5 X 60 cm TSK G 3000 SW column in 6 M guanidine hydrochloride. Their molecular weights estimated by GPC were about 29,000 daltons for the beta subunit and 19,000 daltons for the alpha subunit. Both subunits were examined for SDS-polyacrylamide gel electrophoresis, amino acid analyses and radioimmunoassay, respectively. Neither subunit showed significant binding to the hCG-receptor fraction of rat Leydig cell in vitro. However, the separated subunits easily recombined and recovered about 90% of receptor binding activity of intact hCG. The present results indicate that the GPC technique is an effective, simple and non-denaturing method for the separation of hCG into alpha and beta subunits.
Y
Y
Yuki
R
R
Nishimura
M
M
Mochizuki
D000596:Amino Acids / Q000032:analysis; D006063:Chorionic Gonadotropin / Q000032:analysis* / Q000302:isolation & purification* / Q000652:urine; D018997:Chorionic Gonadotropin, beta Subunit, Human; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015292:Glycoprotein Hormones, alpha Subunit; D006801:Humans; D010446:Peptide Fragments / Q000302:isolation & purification*; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000187:drug effects; D011974:Receptors, LH
D000596:Amino Acids; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D015292:Glycoprotein Hormones, alpha Subunit; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011974:Receptors, LH