Transport of branched-chain amino acids in membrane vesicles of Streptococcus cremoris

J Bacteriol. 1987 Nov;169(11):5193-200. doi: 10.1128/jb.169.11.5193-5200.1987.

Abstract

The kinetics, specificity, and mechanism of branched-chain amino acid transport in Streptococcus cremoris were studied in a membrane system of S. cremoris in which beef heart mitochondrial cytochrome c oxidase was incorporated as a proton motive force (delta p)-generating system. Influx of L-leucine, L-isoleucine, and L-valine can occur via a common transport system which is highly selective for the L-isomers of branched chain amino acids and analogs. The pH dependency of the kinetic constants of delta p-driven L-leucine transport and exchange (counterflow) was determined. The maximal rate of delta p-driven transport of L-leucine (Vmax) increased with increasing internal pH, whereas the affinity constant increased with increasing external pH. The affinity constant for exchange (counterflow) varied in a similar fashion with pH, whereas Vmax was pH independent. Further analysis of the pH dependency of various modes of facilitated diffusion, i.e., efflux, exchange, influx, and counterflow, suggests that H+ and L-leucine binding and release to and from the carrier proceed by an ordered mechanism. A kinetic scheme of the translocation cycle of H+-L-leucine cotransport is suggested.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Branched-Chain / metabolism*
  • Amino Acids, Branched-Chain / pharmacology
  • Animals
  • Biological Transport / drug effects
  • Cattle
  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure
  • Electron Transport Complex IV / metabolism
  • Kinetics
  • Leucine / metabolism
  • Mitochondria, Heart / enzymology
  • Models, Biological
  • Streptococcus / metabolism*

Substances

  • Amino Acids, Branched-Chain
  • Electron Transport Complex IV
  • Leucine