Interface residues that drive allosteric transitions also control the assembly of L-lactate Dehydrogenase

The allosteric enzyme, L-lactate dehydrogenase (LDH), is activated by fructose 1,6-metaphosphate (FBP) to reduce pyruvate to lactate. The molecular details of the FBP-driven transition between the low affinity T-state to the high affinity R-state in LDH, a tetramer composed of identical subunits, are not known. The dynamics of theT→R allosteric transition, investigated using Brownian dynamics (BD) simulations of the Self-Organized Polymer (SOP) model, revealed that coordinated rotations of the subunits drive the T→R transition. We used the structural perturbation method (SPM), which requires only the static structure, to identify the allostery wiring diagram (AWD), a network of residues that transmits signals across the tetramer, as LDH undergoes the T→R transition. Interestingly, the residues that play a major role in the dynamics, which are predominantly localized at the interfaces, coincide with the AWD identified using the SPM. The conformational changes in the T→R transition start from the region near the active site, comprising of helix αC, helix α1/2G, helix α3G and helix α2F, and proceed to other structural units, thus completing the global motion. Brownian dynamics simulations of the tetramer assembly, triggered by a temperature quench from the fully disrupted conformations, show that the bottleneck for assembly is the formation of the correct orientation between the subunits, requiring contacts between the interface residues. Surprisingly, these residues are part of the AWD, which was identified using the SPM. Taken together, our results show that LDH, and perhaps other multi-domain proteins, may have evolved to stabilize distinct states of allosteric enzymes using precisely the same AWD that also controls the functionally relevant allosteric transitions.