RECONSTITUTION OF THE HISTIDINE PERIPLASMIC TRANSPORT-SYSTEM IN MEMBRANE-VESICLES - ENERGY COUPLING AND INTERACTION BETWEEN THE BINDING-PROTEIN AND THE MEMBRANE COMPLEX

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  • 25 March 1989
    • journal article
    • research article
    • Vol. 264  (9) , 5006-5014
Abstract
The periplasmic histidine transport system of Salmonella typhimurium has been reconstituted in isolated right-side-out membrane vesicles. The reconstituted system is entirely dependent on both the periplasmic protein, HisJ, and the membrane-bound complex, composed of proteins HisQ, HisM, and HisP. Transport is also dependent on the presence of ascorbate and phenazine methosulfate, which provide the energy for transport. Ascorbate oxidation generates a proton-motive-force, which allows ATP synthesis. ATP (or a cogenerated molecule) appears to be the immediate energy donor. Dissipation of the proton-motive-force or reduction of the level of ATP by a variety of treatments results in inhibition of transport. Vanadate inhibits transport, indicating that ATP utilization is necessary to energize transport. The interaction between liganded HisJ and the membrane complex has been measured directly: it displays Michaelis-Menten type kinetics, with a K1/2 of .apprx. 65 .mu.M. The significance of this finding in terms of transport prperties of whole cell is discussed.