Recombinant aequorin as a probe for cytosolic free Ca2+ in Escherichia coli.
We describe a novel and simple method for the measurement of bacterial cytosolic free calcium ([Ca2+]i) using recombinant aequorin reconstituted within live bacterial cells. Using this method we have measured the effects of external calcium, complement, phagocytosis and antibiotics on the [Ca2+]i of Escherichia coli. In principle this method should be applicable to any genetically transformable organism and should suffer fewer problems than fluorescent dyes for subcellular calcium measurement.
Knight
M R
MR
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Campbell
A K
AK
Smith
S M
SM
Trewavas
A J
AJ
D000331:Aequorin / Q000031:analogs & derivatives / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001059:Apoproteins / Q000737:chemistry; D002118:Calcium / Q000378:metabolism*; D003165:Complement System Proteins / Q000494:pharmacology; D004926:Escherichia coli / Q000502:physiology*; D006801:Humans; D007093:Imidazoles; D066298:In Vitro Techniques; D049449:Luminescence; D009504:Neutrophils / Q000502:physiology; D011719:Pyrazines; D011994:Recombinant Proteins; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D001059:Apoproteins; D007093:Imidazoles; D011719:Pyrazines; D011994:Recombinant Proteins; C017144:coelenterazine; D000331:Aequorin; D003165:Complement System Proteins; D002118:Calcium