Open AccessPurification of Bovine Kidney and Heart Pyruvate Dehydrogenaseb Phosphatase on Sepharose Derivatized with the Pyruvate Dehydrogenase Complex
Author(s) -
PRATT Mary L.,
MAHER James F.,
ROCHE Thomas E.
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb06690.x
Subject(s) - pyruvate dehydrogenase complex , pyruvate dehydrogenase phosphatase , sepharose , biochemistry , pyruvate dehydrogenase kinase , dihydrolipoyl transacetylase , pyruvate decarboxylation , affinity chromatography , lipoic acid , chemistry , dehydrogenase , branched chain alpha keto acid dehydrogenase complex , acid phosphatase , chromatography , enzyme , antioxidant
Pyruvate dehydrogenaseb phosphatase has been purified to apparent homogeneity from mitochondria1 extracts of both beef heart and beef kidney. An essential step in this three‐step purification is affinity chromatography of a largely purified phosphatase fraction using Sepharose beads to which pyruvate dehydrogenase complex is covalently bound through the lipoic acid residues of the dihydrolipoyl transacetylase component of the complex. The purified phosphatase, which has a native relative molecular mass, M r of about 140000, is composed of two nonidentical subunits of M r 89000 and 49000.