Altering CLC stoichiometry by reducing non-polar side-chains at the dimerization interface.

ABSTRACT: CLC-ec1 is a Cl^-/H⁺ antiporter that forms stable homodimers in lipid bilayers, with a free energy of -10.9 kcal/mol in 2:1 POPE/POPG lipid bilayers. The dimerization interface is formed by four transmembrane helices: H, I, P and Q, that are lined by non-polar side-chains that come in close contact, yet it is unclear as to whether their interactions drive dimerization. To investigate whether non-polar side-chains are required for dimer assembly, we designed a series of constructs where side-chain packing in the dimer state is significantly reduced by making 4-5 alanine substitutions along each helix (H-ala, I-ala, P-ala, Q-ala). All constructs are functional and three purify as stable dimers in detergent micelles despite the removal of significant side-chain interactio

SUBMITTER: Mersch K 

PROVIDER: S-EPMC8734038 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature