Modification of Rhodospirillum rubrum ribulose bisphosphate carboxylase with pyridoxal phosphate. 1. Identification of a lysyl residue at the active site.
Ribulose 1,5-bisphosphate carboxylase isolated from Rhodospirillum rubrum was strongly inhibited by low concentrations of pyridoxal 5'-phosphate. Activity was protected by the substrate ribulose bisphosphate and to a lesser extent by other phosphorylated compounds. Pyridoxal phosphate inhibition was enhanced in the presence of magnesium and bicarbonate, but not in the presence of either compound alone. Concomitant with inhibition of enzyme activity, pyridoxal phosphate forms a Schiff base with the enzyme which is reversible upon dialysis and reducible with sodium borohydride. Subsequent to reduction of the Schiff base with tritiated sodium borohydride, tritiated N6-pyridoxyllysine could be identified in the acid hydrolysate of the enzyme. Only small amounts of this compound were present when the reduction was performed in the presence of carboxyribitol bisphosphate, an analogue of the intermediate formed during the carboxylation reaction. Therefore, it is concluded that pyridoxal phosphate modifies a lysyl residue close to or at the active site of ribulose bisphosphate carboxylase.
D001639:Bicarbonates / Q000494:pharmacology; D001665:Binding Sites; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors*; D002712:Chlorides / Q000494:pharmacology; D006600:Hexosephosphates / Q000494:pharmacology; D008239:Lysine; D008274:Magnesium / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D010428:Pentosephosphates / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000494:pharmacology*; D012247:Rhodospirillum rubrum / Q000201:enzymology; D012273:Ribulose-Bisphosphate Carboxylase / Q000037:antagonists & inhibitors*; D012545:Schiff Bases
D001639:Bicarbonates; D002712:Chlorides; D006600:Hexosephosphates; D010428:Pentosephosphates; D012545:Schiff Bases; D009249:NADP; D011732:Pyridoxal Phosphate; D002262:Carboxy-Lyases; D012273:Ribulose-Bisphosphate Carboxylase; D008274:Magnesium; D008239:Lysine