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Identification of gamma‐glutamyltransferase in rat liver plasma membranes after two‐dimensional electrophoresis
Author(s) -
RahimiPour Ali,
WellmanBednawska Maria,
Galteau MarieMadeleine,
Artur Yves,
Siest GÉRard
Publication year - 1986
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150070206
Subject(s) - isoelectric focusing , molecular mass , antiserum , gel electrophoresis , microbiology and biotechnology , protein subunit , silver stain , chemistry , isoelectric point , kidney , biochemistry , antibody , gamma glutamyltransferase , sodium dodecyl sulfate , polyacrylamide gel electrophoresis , enzyme , biology , endocrinology , gene , immunology
Two‐dimensional electrophoresis and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, followed by silver staining or specific immunoblotting analysis, was applied to the characterization of gamma‐glutamyltransferase (GGT, EC 2.3.2.2.) in rat tissues. We confirmed that purified kidney GGT is a dimer composed of two non‐identical subunits with molecular masses of about 50 and 30 kDa. Both the light and the heavy subunits were separated into 6 and 2 protein bands, respectively. Antibody fractions against the 30 and 50 kDa subunits, purified by immunoaffinity from a whole antiserum directed to the rat kidney enzyme, recognized their corresponding subunits and did not cross‐react with each other. Studies of the reactivity of these antibodies towards GGTs from kidney and liver homogenates revealed an intraspecies dissimilarity in the molecular architecture of the kidney and liver GGTs, especially concerning the 30 kDa subunit. Following phenobarbital treatment of animals we observed an increase in immunoreactive GGT, accompanied by the appearance of additional polypeptides with more basic isoelectric points and higher molecular mass.