Evidence for a substrate assisted conformational transformation of glyceraldehyde 3-phosphate dehydrogenase.

Glyceraldehyde 3-phosphate dehydrogenase exhibits half-site reactivity, the structural origin of which is obscure. Thermal inactivation kinetics, employed here as a probe for site-site heterogeneity in solution, show that green gram glyceraldehyde 3-phosphate dehydrogenase (in the absence and presence of phosphate and NAD+) loses activity in two distinct phases, each of which accounts for half of the initial activity. In the presence of substrate, glyceraldehyde 3-phosphate the relative amplitude of the slow phase increases, and at 0.06 mM glyceraldehyde 3-phosphate the time-course of inactivation corresponds to a single exponential decay. The data are consistent with a suggestion that glyceraldehyde 3-phosphate dehydrogenase may exist in two interconvertible conformations of different symmetry characteristics (C2 in equilibrium D2). The lower symmetry conformation (C2) predominates in the apoenzyme and in the presence of phosphate and NAD+. The higher symmetry conformation (D2) is stabilised by glyceraldehyde 3-phosphate.