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. 1974 Dec;71(12):5032–5036. doi: 10.1073/pnas.71.12.5032

Determination of the Absolute Number of Escherichia coli Membrane Vesicles That Catalyze Active Transport*,†

S A Short 1, H R Kaback 1,‡, G Kaczorowski §, J Fisher §, C T Walsh §, S C Silverstein
PMCID: PMC434034  PMID: 4612538

Abstract

Transport of vinylglycolate (2-hydroxy-3-butenoic acid) via the lactate transport system is the limiting step for covalent labeling of membrane vesicles prepared from E. coli ML 308-225. Thus, the rate and extent of vinylglycolate labeling is stimulated about 10-fold by ascorbate-phenazine methosulfate, and stimulation is abolished by 2,4-dinitrophenol and by phospholipase treatment, neither of which affect the rate of vinylglycolate oxidation. [3H]Vinylglycolate of high specific activity has been prepared, and vesicles have been labeled with this compound in the presence of ascorbate-phenazine methosulfate. Examination of these preparations by high resolution radioautography in the electron microscope demonstrates that virtually all of the vesicles are labeled. The experiments provide a strong indication that most, if not all, of the membrane vesicles in these preparations catalyze active transport.

Keywords: vinylglycolate, 2-hydroxy-3-butenoate, D-lactate, radioautography, electron microscopy

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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