Referenced in: none

Automatically associated channels: Kv10.1

Title: Ca2+ selectivity of a chemically modified OmpF with reduced pore volume.

Authors: Henk Miedema, Maarten Vrouenraets, Jenny Wierenga, Dirk Gillespie, Bob Eisenberg, Wim Meijberg, Wolfgang Nonner

Journal, date & volume: Biophys. J., 2006 Dec 15 , 91, 4392-400

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/16997866

Abstract
We studied an E. coli OmpF mutant (LECE) containing both an EEEE-like locus, typical of Ca(2+) channels, and an accessible and reactive cysteine. After chemical modification with the cysteine-specific, negatively charged (-1e) reagents MTSES or glutathione, this LECE mutant was tested for Ca(2+) versus alkali metal selectivity. Selectivity was measured by conductance and zero-current potential. Conductance measurements showed that glutathione-modified LECE had reduced conductance at Ca(2+) mole fractions <10(-3). MTSES-modified LECE did not. Apparently, the LECE protein is (somehow) a better Ca(2+) chelator after modification with the larger glutathione. Zero-current potential measurements revealed a Ca(2+) versus monovalent cation selectivity that was highest in the presence of Li(+) and lowest in the presence of Cs(+). Our data clearly show that after the binding of Ca(2+) the LECE pore (even with the bulky glutathione present) is spacious enough to allow monovalent cations to pass. Theoretical computations based on density functional theory combined with Poisson-Nernst-Planck theory and a reduced pore model suggest a functional separation of ionic pathways in the pore, one that is specific for small and highly charged ions, and one that accepts preferentially large ions, such as Cs(+).