Interrelations of ATP synthesis and proton handling in ischaemically exercising human forearm muscle studied by 31P magnetic resonance spectroscopy.

1. In ischaemic exercise ATP is supplied only by glycogenolysis and net splitting of phosphocreatine (PCr). Furthermore, 'proton balance' involves only glycolytic lactate/H+ generation and net H+ 'consumption' by PCr splitting. This work examines the interplay between these, metabolic regulation and the creatine kinase equilibrium. 2. Nine male subjects (age 25-45 years) performed finger flexion (7 % maximal voluntary contraction at 0.67 Hz) under cuff ischaemia. 31P magnetic resonance spectra were acquired from finger flexor muscle in a 4.7 T magnet using a 5 cm surface coil. 3. Initial PCr depletion rate estimates total ATP turnover rate; glycolytic ATP synthesis was obtained from this and changes in [PCr], and then used to obtain flux through 'distal' glycolysis (phosphofructokinase and beyond) to lactate; 'proximal' flux (through phosphorylase) was obtained from this and changes in [phosphomonoester]. Total H+ load (lactate load less H+ consumption) was used to estimate cytosolic buffer capacity (beta). 4. Glycolytic ATP synthesis increased from near zero while PCr splitting declined. Net H+ load was approximately linear with pH, suggesting beta = 20 mmol x l(-1) (pH unit)(-1) at rest, increasing as pH falls. 5. Relationships between glycolytic rate and changes in [PCr] (i.e. the time-integrated mismatch between ATP use and production), and thus also [P(i)] (substrate for phosphorylase), suggest that increase in glycolysis is due partly to 'open-loop' Ca2+-dependent conversion of phosphorylase b to a, and partly to the 'closed loop' increase in P(i) consequent on net PCr splitting. 6. The 'settings' of these mechanisms have a strong influence on changes in pH and metabolite concentrations.