A high-throughput screen for MscL channel activity and mutational phenotyping

A novel fluorescence-based screen for bacterial mechanosensitive ion-channel activity has been developed. This assay is capable of clearly distinguishing the previously observed gain of function and loss of function phenotypes for the Escherichia coli mechanosensitive channel of large conductance (E...

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Bibliographic Details
Published inBiochimica et biophysica acta Vol. 1514; no. 2; pp. 165 - 169
Main Authors Maurer, Joshua A, Dougherty, Dennis A
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.10.2001
Subjects
Cell Survival
Escherichia coli - genetics
Escherichia coli - growth & development
Escherichia coli Proteins
Fluorescence
Gain of function
Ion channel
Ion Channels - analysis
Ion Channels - genetics
Loss of function
Mechanosensation
MscL
Mutation
Osmotic Pressure
Phenotype
Propidium
Ec, Escherichia coli
Fluorescence
Loss of function
MscL
GOF, gain of function
Ion channel
Mechanosensation
MscS, mechanosensitive channel of small conductance
LOF, loss of function
Tb, Mycobacterium tuberculosis
HOEM, high osmolyte enhanced M9
Gain of function
MscL, mechanosensitve channel of large conductance
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Summary:A novel fluorescence-based screen for bacterial mechanosensitive ion-channel activity has been developed. This assay is capable of clearly distinguishing the previously observed gain of function and loss of function phenotypes for the Escherichia coli mechanosensitive channel of large conductance (Ec-MscL). The method modifies Molecular Probes’ Live/Dead® BacLight™ bacterial viability assay to monitor MscL channel activity as a function of bacterial survival from osmotic downshock.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:0005-2736
0006-3002
1879-2642
DOI:10.1016/S0005-2736(01)00390-X