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PMID: 1577684 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The efflux of a fluorescent probe is catalyzed by an ATP-driven extrusion system in Lactococcus lactis.

Journal of bacteriology ·Vol. 174 ·No. 10 ·1992-05-00 ·Pages 3118-24

Molenaar D, Bolhuis H, Abee T, Poolman B, Konings WN

Abstract

Many bacteria, both gram positive and gram negative, extrude in an energy-dependent manner the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5[and -6]-carboxyfluorescein (BCECF) (D. Molenaar, T. Abee, and W. N. Konings, Biochim. Biophys. Acta 1115:75-83, 1991). This efflux was studied in detail in Lactococcus lactis, and several indications that a transport system is involved were found. This transport system is most likely driven by ATP or a related compound. The evidence is that BCECF extrusion (i) occurs against a BCECF gradient, (ii) is strictly correlated with ATP concentration and not with the proton motive force, and (iii) is inhibited by vanadate and to a lesser extent by N,N'-dicyclohexylcarbodiimide. Most convincingly, a UV mutant with a strongly reduced efflux rate was isolated. Such a mutant was isolated from a BCECF-loaded and lactose-energized population by selection of highly fluorescent cells in a flow cytometer-cell sorter. The physiological function of this extrusion system is unknown, but its characteristics classify it among the traffic ATPases.

MeSH Terms
Adenosine Triphosphate/metabolism Biological Transport, Active/drug effects Dicyclohexylcarbodiimide/pharmacology Drug Resistance, Microbial Flow Cytometry Fluoresceins/metabolism Hot Temperature Lactococcus lactis/drug effects,metabolism Lactose/metabolism Mutation Vanadates/pharmacology
Chemicals
Fluoresceins Vanadates Dicyclohexylcarbodiimide 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein Adenosine Triphosphate Lactose
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Molenaar D
Laboratory for Microbiology, University of Groningen, Haren, The Netherlands.
Bolhuis H
Abee T
Poolman B
Konings W N
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.10.3118-3124.1992
Published
1992-05-00
Pages
3118-24
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205976
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.174.10.3118-3124.1992 PMC Full Text View journal details
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