Distinct metal cofactor-induced conformational states in the NAD-specific malic enzyme of Escherichia coli as revealed by proteolysis studies.
Evidence is presented for the existence of altered ligand-stabilized conformational states of the NAD-specific malic enzyme (L-malate:NAD+ oxidoreductase (oxaloacetate-decarboxylating), EC 1.1.1.38), of Escherichia coli in the presence of Mg2+ and Mn2+, as identified by their susceptibilities to proteolysis. The rate of tryptic digestion of the enzyme is significantly decreased in the Mg2+-form of the enzyme when the product, NADH, or the allosteric effectors, coenzyme A and aspartate, are present in the digestion mixture. In contrast, little difference in the rate of tryptic digestion is observed in the degree of protection of the enzyme by the two metal cofactors, either alone, or in the presence of the substrates, malate and NAD. The results are consistent with the previously proposed hypothesis of Milne and Cook (Biochemistry 18, (1979) 3604-3610) that Mg2+ and Mn2+ stabilize two distinct conformational states of the enzyme. The results are discussed in relation to the altered kinetic response of the enzyme to substrates and effectors in the presence of the two metal cofactors.
D000494:Allosteric Regulation; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D008024:Ligands; D008274:Magnesium / Q000378:metabolism; D008291:Malate Dehydrogenase; D008345:Manganese / Q000378:metabolism; D009243:NAD; D011487:Protein Conformation; D014357:Trypsin
D008024:Ligands; D009243:NAD; D008345:Manganese; D008291:Malate Dehydrogenase; D014357:Trypsin; D008274:Magnesium