| Abstract: | Pyridoxal(5')diphospho(1)-alpha-D-glucose has been tested as an inhibitor of native glycogen phosphorylases a and b. Its inhibition patterns with respect to substrate, glucose 1-phosphate, and activator, adenosine monophosphate, show it to be a potent (Ki = 40 microM) R-state inhibitor of phosphorylase b, mimicking the binding of glucose-1-phosphate, and, as predicted for an R-state inhibitor, its binding to AMP-activated phosphorylase a is even tighter (Ki = 10 microM). Moreover, it is demonstrated that its binding does not involve covalent imine formation from the pyridoxal aldehyde to an active-site lysine residue. It thus represents the tightest binding R-state inhibitor reported to date, and a 31P NMR study of the effects of binding of this inhibitor upon 31P resonances for the coenzyme phosphate and that of the nucleotide activator is presented. Results obtained are essentially identical to those obtained previously using glucose cyclic 1,2-phosphate, corroborating the previous conclusions. A rationale for the tightness of the binding is presented, as are other possible uses of this compound in studies on glycogen phosphorylase and other similar enzymes. |
