Making Monomeric Aquaporin Z by Disrupting the Hydrophobic Tetramer Interface.

The assembly of integral membrane proteins depends on the packing of hydrophobic interfaces. The forces driving this packing remain unclear. In this study, we have investigated the effect of mutations in these hydrophobic interfaces on the structure and function of the tetrameric water channel aquaporin Z (AqpZ). Among the variants, we have constructed several fail to form tetramers and are monomeric. In particular, both of the mutants which are expected to create interfacial cavities become monomeric. Furthermore, one of the mutations can be compensated by a second-site mutation. We suggest that the constraints imposed by the nature of the lipid solvent result in interfaces that respond differently to modifications of residues. Specifically, the large size and complex conformations of lipid molecules are unable to fill small interfacial holes. Further, we observe in AqpZ that there is a link between the oligomeric state and the water channel activity. This despite the robustness of both protein folding and topology, both of which remain unchanged by the mutations we introduce. We propose that this linkage may result from the specific modes of structural flexibility in the monomeric protein.