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Purification and characterization of guinea‐pig epidermal acid phosphatase
Author(s) -
MIYAGAWA T.,
ANAI M.,
URABE H.
Publication year - 1977
Publication title -
british journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.304
H-Index - 179
eISSN - 1365-2133
pISSN - 0007-0963
DOI - 10.1111/j.1365-2133.1977.tb06135.x
Subject(s) - sephadex , isoelectric point , size exclusion chromatography , chemistry , chromatography , acid phosphatase , enzyme , divalent , phosphate , biochemistry , deoxyribonucleoside , hydrolysis , isoelectric focusing , organic chemistry
SUMMARY Guinea‐pig epidermal acid phosphatase has been purified approximately 120‐fold by a procedure including acid treatment, CM‐cellulose and DEAE‐cellulose chromatography, and gel filtration on Sephadex G‐100. The enzyme had a pH optimum at 5.0 and the optimal temperature for activity was approximately 50 C. The enzyme was not activated by divalent cations or 2‐mercaptoethanol, but it was inhibited by p‐chloromercuribenzoate and by fluoride. The km value for p‐nitrophenyl phosphate was 1.31 × 10 −4 M, the molecular weight was about 73,000 as determined by Sephadex G‐100 gel filtration and the isoelectric point was 6.1 The enzyme hydrolyzed deoxyribonucleoside monophosphates to deoxyribonucleosides.