Targeting of aequorin for calcium monitoring in intracellular compartments.
We have recently developed a new method for monitoring Ca2+ concentrations in defined cell compartments. The cDNA encoding the Ca(2+)-sensitive photoprotein aequorin has been modified in order to include specific targeting sequences and expressed in eukaryotic cells; the recombinant protein, specifically located inside the cells, has allowed the direct study of mitochondrial and nuclear Ca2+ concentrations in living cells. The principles, and the application, of this new methodology are discussed in this article.
Brini
M
M
Department of Biomedical Sciences, University of Padova, Italy.
Pasti
L
L
Bastianutto
C
C
Murgia
M
M
Pozzan
T
T
Rizzuto
R
R
D000331:Aequorin / Q000378:metabolism*; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002999:Clone Cells; D016257:Fura-2; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014162:Transfection
D011994:Recombinant Proteins; D000331:Aequorin; D002118:Calcium; D016257:Fura-2