Does pyridoxal 5'-phosphate function in glycogen phosphorylase as an electrophilic or a general acid catalyst?

• Published in: Biochemistry (Easton) 1984-11, Vol.23 (24), p.5853-5861
• Main Authors: Klein, Helmut W, Im, Mie J, Palm, Dieter, Helmreich, Ernst J. M
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Catalysis; Diphosphates - metabolism; Enzyme Activation; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Glucosephosphates - metabolism; Hydrogen-Ion Concentration; Ions; Magnetic Resonance Spectroscopy; muscles; Muscles - enzymology; N.M.R; phosphorylase; Phosphorylase b - metabolism; Phosphorylases - metabolism; Pyridoxal - analogs & derivatives; Pyridoxal - metabolism; pyridoxal 5'-phosphate; Pyridoxal Phosphate - analogs & derivatives; Pyridoxal Phosphate - metabolism; Rabbits; Sugar Phosphates; Transferases
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00319a027

alpha-D-Glucose 1-diphosphate interacts with pyridoxal-reconstituted rabbit muscle phosphorylase b activated by AMP (AMP-S). Under these conditions, the glucose moiety of alpha-D-[14C]glucose 1-diphosphate is transferred to limit dextrin forming alpha(1---4) glycosidic bonds and simultaneously releasing pyrophosphate as shown by 31P NMR spectroscopy. Thus, specific structural requirements invoked to explain the reactions of pyridoxal(5')diphospho(1)-alpha-D-glucose need not to be assumed in the case of the reactions of alpha-D-glucose 1-diphosphate. Dianions isomorphous to phosphate activate pyridoxal phosphorylase regardless of their pK values while the same anions, when bound covalently to pyridoxal, are inactive. Thus, anions bound noncovalently to pyridoxal phosphorylase act differently than anions linked covalently to pyridoxal, such as the 5'-phosphate group of pyridoxal 5'-phosphate, which is postulated to be part of a proton donor-acceptor pathway. The reaction of 2,6-anhydro-1-deoxy-D-gluco-hept-1-enitol (heptenitol) with phosphorylase yields, in the presence of orthophosphate as a glycosyl acceptor, 1-deoxy-D-gluco-heptulose 2-phosphate (heptulose-2-P). This sugar phosphate is unreactive but a potent competitive inhibitor for rabbit muscle phosphorylase b and potato phosphorylase with respect to alpha-D-glucose 1-phosphate: Ki = 14 X 10(-6) M and 1.9 X 10(-6) M, respectively.