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PURIFICATION OF PHOSPHATE‐DEPENDENT PIG BRAIN GLUTAMINASE
Author(s) -
Svenneby G.,
Torgner I. Aa.,
Kvamme E.
Publication year - 1973
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.1471-4159.1973.tb00090.x
Subject(s) - chromatography , chemistry , isoelectric point , tris , centrifugation , polyacrylamide gel electrophoresis , phosphate , sodium , acetone , sedimentation equilibrium , gel electrophoresis , extraction (chemistry) , fractionation , electrophoresis , polyacrylamide , isoelectric focusing , biochemistry , enzyme , ultracentrifuge , organic chemistry , polymer chemistry
— A procedure for preparing highly purified phosphate‐activated glutaminase (EC 3.5.1.2, L‐glutamine amidohydrolase) from pig brain is described. The main steps consist of extraction with acetone, followed by sodium sulphate fractionation of the solubilized acetone powder. Thereafter, solubilization by dialysis against a buffer containing tris‐HC1, mercaptoethanol, and EDTA, followed by precipitation with phosphate‐borate, is repeated twice. The final preparation contains no impurities which can be detected by polyacrylamide gel electrophoresis, isoelectric focusing, and sedimentation equilibrium centrifugation. By the latter method, molecular weight is determined to be 187,000. By polyacrylamide gel electrophoresis in sodium dodecyl sulphate, one protein band with molecular weight 64,000 is found.