Use of small fluorescent molecules to monitor channel activity.

Abstract:

:The Mechanosensitive channel of Large conductance (MscL) allows bacteria to rapidly adapt to changing environmental conditions such as osmolarity. The MscL channel opens in response to increases in membrane tension, which allows for the efflux of cytoplasmic constituents. Here we describe the cloning and expression of Salmonella typhimurium MscL (St-MscL). The amino acid sequence encoding for this MscL exhibits a high degree of similarity to Escherichia coli MscL (Eco-MscL). Using a fluorescence efflux assay, we demonstrate that efflux through the MscL channel during hypoosmotic shock can be monitored using endogenously produced fluorophores. These fluorophores are synthesized by a cotransformed gene, cobA. In addition, we observe that thermal stimulation, i.e., heat shock, can induce efflux through MscL.

authors

Jones SE,Naik RR,Stone MO

doi

10.1006/bbrc.2000.3921

subject

Has Abstract

pub_date

2000-12-09 00:00:00

pages

208-12

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(00)93921-4

journal_volume

279

pub_type

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