Inhibition of the Cardiac Sarcoplasmic Reticulum Ca2+-ATPase by Glucose 6-Phosphate is Ca2+ Dependent
Defects in the structure or function of the cardiac sarcoplasmic reticulum (CSR) Ca2+-ATPase presumably contribute to the Ca2+ imbalance in the diabetic myocardium. The susceptibility to nonenzymatic protein glycation by glucose metabolites is suggested due to the relatively high percent of target lysines and arginines (approaching 15 mol%) at the ATP binding and phosphorylation domains. Brief incubations (15 min) of CSR microsomes at 24°C in the presence of 5.0 mM glucose 6-phosphate (G1c6P) inhibited Ca2+-dependent ATPase maximal activity relative to controls. Inhibition was only observed when incubations contained 0.1 mM CaCl2 (1.86 μmol ATP hydrolyzed · mg−1 · min−1, +Glc6P versus 2.78, control). Nonconvergent regression lines drawn from maximal velocities as a function of CSR microsome concentration indicate an irreversible mechanism of inhibition which is supported by an observed depletion in CSR amine content (2.98 μmol -NH2 groups/mg microsomal protein, +Gl6P versus 3.34, control). Glucose 6-phosphate (5.0mM) in Ca2+-free incubations (plus 0.1 mM EGTA) had no affect on either enzyme activity or total amine content. These data suggest that the E1 but not the E2 conformation of the CSR Ca2+-ATPase is susceptible to Glc6P-mediated modification resulting in diminished maximal Ca2+-dependent ATPase activity.
Kraft A, Cassetta M, Seidler NW. Inhibition of the Cardiac Sarcoplasmic Reticulum Ca2+-ATPase by Glucose 6-Phosphate is Ca2+ Dependent. Life Sciences. 1998; 62(4). doi: 10.1016/s0024-3205(97)01109-0.