Purification and characterization of recombinant human activin B.
Recombinant human activin B has been isolated to more than 95% purity from a mammalian kidney cell line. Activin B is a covalently-linked homodimer with an apparent molecular mass of 25.9 kDa (unreduced) and 15.2 kDa (reduced) as determined by SDS-polyacrylamide-gel electrophoresis. On gel filtration in 6 M guanidine hydrochloride, activin B chromatographs with an apparent molecular mass of 11 kDa, whether reduced or not. The amino-terminal sequence of the purified protein is consistent with the expected sequence derived from the beta subunit of inhibin B. The amino acid composition of the purified molecule agrees with the expected theoretical composition of the beta subunit of inhibin B. Activin B has an apparent pI of 4.6 as determined by isoelectric focusing in 6 M urea and 4.7 as determined by chromatofocusing in 6 M urea. The extinction coefficient is 1.8.
Schmelzer
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CH
Genentech, Inc., Recovery Process R&D, South San Francisco, CA 94080.
Burton
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LE
Tamony
C M
CM
Schwall
R H
RH
Mason
A J
AJ
Liegeois
N
N
D028341:Activins; D000595:Amino Acid Sequence; D002460:Cell Line; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D007265:Inhibins / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D007668:Kidney; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides; D010455:Peptides / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification
D006146:Guanidines; D046911:Macromolecular Substances; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins; C044147:activin B; D028341:Activins; C048617:myelopeptides; D007265:Inhibins; D019791:Guanidine