CLC Cl /H+ transporters constrained by covalent cross-linking.

ABSTRACT: CLC Cl(-)/H(+) exchangers are homodimers with Cl(-)-binding and H(+)-coupling residues contained within each subunit. It is not known whether the transport mechanism requires conformational rearrangement between subunits or whether each subunit operates as a separate exchanger. We designed various cysteine substitution mutants on a cysteine-less background of CLC-ec1, a bacterial CLC exchanger of known structure, with the aim of covalently linking the subunits. The constructs were cross-linked in air or with exogenous oxidant, and the cross-linked proteins were reconstituted to assess their function. In addition to conventional disulfides, a cysteine-lysine cross-bridge was formed with I(2) as an oxidant. The constructs, all of which contained one, two, or four cross-bridges, were function

SUBMITTER: Nguitragool W 

PROVIDER: S-EPMC2409211 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature