Open AccessMechanism and energetics of a citrate‐transport system of Klebsiella pneumoniae
Author(s) -
REST Michel E.,
ABEE Tjakko,
MOLENAAR Douwe,
KONINGS Wil N.
Publication year - 1991
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1991.tb15677.x
Subject(s) - symporter , klebsiella pneumoniae , chemistry , citric acid , membrane , escherichia coli , membrane transport , biophysics , biochemistry , transporter , biology , gene
The citrate‐transport determinant of plasmid pES 1 from Klebsiella pneumoniae [Schwarz, E. & Oesterhelt, D. (1985) EM BO J. 4 , 1599–1603] has been subcloned in Escherichia coli DH1. Uptake of citrate in E. coli membrane vesicles via this uptake system is an electrogenic process, although the pH gradient is the main driving force for citrate uptake. The rate of citrate uptake, driven by artificially imposed ion‐gradients, is high in the presence of an artificial ΔpH and low in the presence of an artificial ΔΨ. Citrate transport does not depend on the presence of Na + or Mg 2+ as has been observed for other citrate‐transport systems. Citrate has three p K values: 3.14, 4.77 and 5.40. Citrate forms a stable complex with Mg 2+ with a stability constant of 3.2. Kinetic parameters and calculations of the different citrate (Cit) species at a given pH, indicate that the HCit 2− is the species transported and that transport occurs in symport with three protons. This citrate‐transport system is thus a unique example of a 3H + /solute symport system.