Simultaneous detection of Ca2+ and diacylglycerol signaling in living cells.
Phospholipase C produces two second messengers--diacylglycerol (DAG), which remains in the membrane, and inositol triphosphate (IP(3)), which triggers the release of calcium ions (Ca(2+)) from intracellular stores. Genetically encoded sensors based on a single circularly permuted fluorescent protein (FP) are robust tools for studying intracellular Ca(2+) dynamics. We have developed a robust sensor for DAG based on a circularly permuted green FP that can be co-imaged with the red fluorescent Ca(2+) sensor R-GECO for simultaneous measurement of both second messengers.
Montana Molecular, Bozeman, Montana, United States of America.
Paul
P
Tewson
Mara
M
Westenberg
Yongxin
Y
Zhao
Robert E
RE
Campbell
Anne Marie
AM
Quinn
Thomas E
TE
Hughes
D015374:Biosensing Techniques / Q000379:methods*; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival; D004075:Diglycerides / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D051745:Protein Kinase C-delta / Q000737:chemistry / Q000378:metabolism; D015290:Second Messenger Systems; D013997:Time Factors
D004075:Diglycerides; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D051745:Protein Kinase C-delta; D002118:Calcium