4.6 Review Book Chapter

It's Better To Be Lucky Than Smart

Journal

ANNUAL REVIEW OF BIOCHEMISTRY, VOL 90, 2021
Volume 90, Issue -, Pages 1-29

Publisher

ANNUAL REVIEWS
DOI: 10.1146/annurev-biochem-011520-105008

Keywords

chemiosmosis; proton electrochemical gradient; membrane vesicles; lactose permease; proton symport; site-directed mutagenesis; autobiography

Funding

  1. National Heart Institute
  2. Roche Institute of Molecular Biology
  3. Howard Hughes Medical Institute
  4. National Institutes of Health
  5. National Science Foundation

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Bacterial cytoplasmic membrane vesicles serve as a unique experimental system for studying active transport, providing a clear definition of the reactions involved in the process. Experimental observations show that respiratory energy in membrane vesicles from Escherichia coli is primarily converted into work in the form of a solute concentration gradient driven by a proton electrochemical gradient, as proposed by Peter Mitchell's chemiosmotic theory.
Bacterial cytoplasmic membrane vesicles provide a unique experimental system for studying active transport. Vesicles are prepared by lysis of osmotically sensitized cells (i.e., protoplasts or spheroplasts) and comprise osmotically intact, unit-membrane-bound sacs that are approximately 0.5-1.0 mu m in diameter and devoid of internal structure. Their metabolic activities are restricted to those provided by the enzymes of the membrane itself, and each vesicle is functional. The energy source for accumulation of a particular substrate can be determined by studying which compounds or experimental conditions drive solute accumulation, and metabolic conversion of the transported substrate or the energy source is minimal. These properties of the vesicle system constitute a considerable advantage over intact cells, as the system provides clear definition of the reactions involved in the transport process. This discussion is not intended as a general review but is concerned with respiration-dependent active transport in membrane vesicles from Escherichia coli. Emphasis is placed on experimental observations demonstrating that respiratory energy is converted primarily into work in the form of a solute concentration gradient that is driven by a proton electrochemical gradient, as postulated by the chemiosmotic theory of Peter Mitchell.

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