Open AccessThe mitochondrial inner‐membrane anion channel possesses two mercurial‐reactive regulatory sites
Author(s) -
BEAVIS Andrew D.
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb15143.x
Subject(s) - mersalyl , chemistry , divalent , protonophore , biophysics , uniporter , binding site , membrane , inorganic chemistry , biochemistry , stereochemistry , mitochondrion , cytosol , biology , organic chemistry , enzyme
The mitochondrial inner membrane anion channel catalyzes the electrophoretic transport of a wide variety of anions and is inhibited by matrix divalent cations and protons. In this paper, evidence is provided that mersalyl and p ‐chloromercuribenzene‐sulfonate each interact with this uniporter at two distinct sites. Binding to site 1 causes a shift in the pH dependence of transport, characterized by a decrease in the pIC 50 for protons from about 7.8 to about 7.3, and leads to substantial stimulation of transport in the physiological pH range. This effect is not reversed by addition of thiols such as thioglycolate. Binding of mersalyl and p ‐chloromercuribenzenesulfonate to site 2 inhibits the transport of most anions including P i , citrate, malonate, sulfate and ferrocyanide. The transport of Cl − is inhibited about 60% by mersalyl, but is not inhibited by p ‐chloromercuribenzenesulfonate. These data suggest that inhibition is a steric effect dependent on the size of the anion and the size of the R group of the mercurial. This inhibition is reversed by thioglycolate. Dose/response curves indicate that mersalyl binds to site 1 as the dose increased from 7 to 13 nmol/mg, whereas it binds to site 2 as the dose is increased from 10 to 18 nmol/mg. Thus, at certain pH values both stimulatory and inhibitory phases can be seen in the same dose/response curve. It is suggested that these sites may contain thiol groups and that physiological regulators may exist which can effect changes in activity of the inner membrane anion uniporter similar to those exerted by mercurials.