title: Probes of catalytic site cooperativity during catalysis by the chloroplast adenosine triphosphate and the adenosine triphosphate synthase.
abstract: During net nucleoside triphosphate synthesis by chloroplast ATP synthase the extent of water oxygen incorporation into each nucleoside triphosphate released increases with decrease in ADP, GDP or IDP concentration. Likewise, during net ATP hydrolysis by the Mg2+-activated chloroplast ATPase, the extent of water oxygen incorporation into each Pi released increases as the ATP, GTP, or ITP concentration is decreased. However, the concentration ranges in which substrate modulation occurs differs with each nucleotide. Modulation of oxygen exchange during synthesis and hydrolysis of adenine nucleotides, as measured by variation in the extent of water oxygen incorporation into products, occurs below 250 microM. In contrast, guanosine and inosine nucleotides alter the extent of exchange at higher and much wider concentration ranges. Activation of the chloroplast ATPase by either heat or trypsin results in similar catalytic behavior as monitored by ATP modulation of oxygen exchanges during hydrolysis in the presence of Mg2+. More exchange capacity is evident with octylglucoside-activated enzyme at all ATP concentrations. High levels of tentoxin were also found to alter the catalytic exchange parameters resulting in continued water oxygen exchange into Pi released during hydrolysis at high ATP concentrations. Little or no oxygen exchange accompanies ATP hydrolysis in the presence of Ca2+. The [18O]Pi species formed from highly gamma-18O-labeled ATP at lower ATP concentrations gives a distribution as expected if only one catalytic pathway is operative at a given ATP concentration. This and other results support the concept of catalytic cooperativity between alternating sites as explanation for the modulation of oxygen exchange by nucleotide concentration.
mesh_terms: D025181:ATP Synthetase Complexes; D000255:Adenosine Triphosphate / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology*; D004789:Enzyme Activation; D005960:Glucosides / Q000494:pharmacology; D008274:Magnesium / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism*; D010101:Oxygen Consumption; D010456:Peptides, Cyclic / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D005960:Glucosides; D009097:Multienzyme Complexes; D010456:Peptides, Cyclic; C018619:octyl-beta-D-glucoside; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D025181:ATP Synthetase Complexes; D014357:Trypsin; D006180:Proton-Translocating ATPases; C003384:tentoxin; D008274:Magnesium; D002118:Calcium