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Large‐Scale, One‐Step Purification of Oxidized and Reduced Forms of Bovine Brain S100b Protein by HPLC
Author(s) -
Mely Y.,
Gérard D.
Publication year - 1988
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1988.tb02976.x
Subject(s) - high performance liquid chromatography , chemistry , chromatography , scale (ratio) , biochemistry , quantum mechanics , physics
A rapid and simple method, using a reverse‐phase column in a HPLC system, has been developed to purify high yields of both oxidized and reduced S100b proteins from a bovine brain S100 protein mixture. The final proteins were characterized by amino acid analysis, sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, and ab‐sorbance and fluorescence spectroscopy. Both S100b subtypes appeared highly purified and differed only by their oxidation state: all four cysteinyl sulfhydryl groups were free in reduced S100b protein whereas two of them gave disulfide bridges in oxidized S100b protein. The stability of the oxidation state of the two isolated subtypes suggests that the two forms were not in rapid equilibrium and probably coexisted in vivo.