A Novel Method for Continuous Determination of the Intracellular pH in Bacteria with the Internally Conjugated Fluorescent Probe 5 (and 6-)-Carboxyfluorescein Succinimidyl Ester.

Applied and Environmental Microbiology
P BreeuwerT Abee

Abstract

A novel method based on the intracellular conjugation of the fluorescent probe 5 (and 6-)-carboxyfluorescein succinimidyl ester (cFSE) was developed to determine the intracellular pH of bacteria. cFSE can be taken up by bacteria in the form of its diacetate ester, 5 (and 6-)-carboxyfluorescein diacetate succinimidyl ester, which is subsequently hydrolyzed by esterases to cFSE in the cytoplasm. When Lactococcus lactis cells were permeabilized with ethanol, a significant proportion of cFSE was retained in the cells, which indicated that cFSE was bound intracellularly. Unbound probe could be conveniently extruded by a short incubation of the cells in the presence of a fermentable sugar, most likely by exploiting an active transport system. Such a transport system for cFSE was identified in L. lactis, Listeria innocua, and Bacillus subtilis. The intracellular pH in bacteria can be determined from the ratio of the fluorescence signal at the pH-sensitive wavelength (490 nm) and the fluorescence signal at the pH-insensitive wavelength (440 nm). This cFSE ratio method significantly reduced problems due to the efflux of fluorescent probe from the cells during the measurement. Moreover, the method described was successfully used to deter...Continue Reading

References

Dec 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·J M SalhanyS Ogawa
Jul 1, 1992·Applied and Environmental Microbiology·M E BrunoT J Montville
Nov 14, 1991·Biochimica Et Biophysica Acta·D MolenaarW N Konings
Oct 15, 1990·Biochemical and Biophysical Research Communications·C N AllenB H Hirst
May 15, 1990·Analytical Biochemistry·M NedergaardW Pulsinelli
Jan 1, 1985·Annual Review of Microbiology·E R Kashket
Dec 1, 1987·Microbiological Reviews·B PoolmanW N Konings
Jul 1, 1986·Analytical Biochemistry·M L GraberE Pastoriza-Munoz
Dec 1, 1985·Microbiological Reviews·I R Booth
Dec 1, 1985·The Journal of General Physiology·J R Chaillet, W F Boron
Oct 1, 1982·The Journal of Cell Biology·T J RinkT Pozzan
Apr 1, 1994·Current Eye Research·M Kondo, M Araie
Jun 1, 1994·Pflügers Archiv : European journal of physiology·N OpitzH Acker
May 2, 1994·Journal of Immunological Methods·A B Lyons, C R Parish

❮ Previous
Next ❯

Citations

Feb 7, 2003·Applied and Environmental Microbiology·Michael G Gänzle, Rudi F Vogel
Jul 6, 2006·Applied and Environmental Microbiology·Sampo J LahtinenSeppo J Salminen
Jul 3, 2007·Applied and Environmental Microbiology·Juan ZhangJian Chen
Mar 10, 2009·Applied and Environmental Microbiology·Jean-Paul MeijnenHarald J Ruijssenaars
Jan 13, 2011·The EMBO Journal·Muhammet E GündoğduLeendert W Hamoen
Jan 1, 2013·Applied and Environmental Microbiology·Dawn E HolmesDerek R Lovley
Jan 8, 2013·Applied Microbiology and Biotechnology·Chongde WuJian Chen
Apr 9, 2014·Antimicrobial Agents and Chemotherapy·Michaël L CartronSimon J Foster
Oct 7, 2014·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Gabriele V M Gabriel, Vadim R Viviani
Sep 9, 2006·Environmental Microbiology·Borja SánchezAbelardo Margolles
Aug 28, 2007·Journal of Bacteriology·Mark A StrauchYoann Le Breton
Sep 19, 2008·Applied Microbiology and Biotechnology·Aline RaultCatherine Béal
Aug 31, 2010·Applied and Environmental Microbiology·Eduardo SánchezNorma Heredia
Feb 20, 2009·Journal of Applied Microbiology·M G Da Silveira, T Abee
Sep 12, 2007·Therapeutic Apheresis and Dialysis : Official Peer-reviewed Journal of the International Society for Apheresis, the Japanese Society for Apheresis, the Japanese Society for Dialysis Therapy·Tomonori ShimakitaHideaki Matsuoka
Mar 31, 2011·Biotechnology and Bioengineering·Stefano GiulittiSara Spilimbergo
Jan 19, 2010·Nature Chemical Biology·Yongfang ZhaoJonathan A Javitch
Jan 23, 2014·Foodborne Pathogens and Disease·Guanghui LiXiaodong Xia

❮ Previous
Next ❯

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.