[Phosphorylation of glyceraldehyde-3-phosphate dehydrogenase].
Phosphorylation of D-glyceraldehyde-3-phosphate dehydrogenase (GPDH) by Ca2+/phospholipid- and Ca2+/calmodulin-dependent protein kinases was shown to take place in rabbit skeletal muscle and brain extracts. The kinases could be "picked up" from the extract, using GPDH immobilized on CNBr-activated Sepharose 4B as an affinity adsorbent. Washing of the column with GPDH solutions resulted in elution of the protein kinases; the same effect was observed when anti-GPDH antibodies were used. The most effective elution took place under the conditions favouring the dissociation of the immobilized GPDH into dimers. Based on these findings, a method for purification of Ca2+/calmodulin-dependent protein kinase has been elaborated, which includes chromatography on phenyl-Sepharose to separate the kinase from GPDH. The susceptibility of GPDH to phosphorylation by tissue protein kinases was confirmed by analyses of GPDH preparations purified from rabbit muscle for endogenous phosphate content: 0.7-1.5 moles of covalently bound phosphate were found per mole of the enzyme.
E A
EA
Sergienko
A A
AA
Ermakova
V I
VI
Muronets
N K
NK
Nagradova
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology; D009943:Organophosphorus Compounds / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011817:Rabbits
D004800:Enzymes, Immobilized; D009943:Organophosphorus Compounds; D000244:Adenosine Diphosphate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases