TY - JOUR
T1 - Synthesis and two-photon photolysis of 6-(ortho-nitroveratryl)-caged IP3 in living cells
AU - Kantevari, Srinivas
AU - Hoang, Caroline J.
AU - Ogrodnik, Jakob
AU - Egger, Marcel
AU - Niggli, Ernst
AU - Ellis-Davies, Graham C.R.
PY - 2006/1
Y1 - 2006/1
N2 - The synthesis of a photolabile derivative of inositol-1,4,5-trisphosphate (IP3) is described. This new caged second messenger (6-ortho-nitroveratryl)-IP3 (6-NV-IP3) has an extinction coefficient of 5000M-1 cm-1 at 350 nm, and a quantum yield of photolysis of 0.12. Therefore, 6-NV-IP3 is photolyzed with UV light about three times more efficiently than the widely used P 4(5)-1-(2-nitrophenyl)ethyl-caged IP3 (NPE-IP 3). 6-NV-IP3 has a two-photon cross-section of about 0.035 GM at 730 nm. This absorbance is sufficiently large for effective two-photon excitation in living cells at modest power levels. Using near-IR light (5 mW, 710 nm, 80 MHz, pulse-width 70 fs), we produced focal bursts of IP3 in HeLa cells, as revealed by laser-scanning confocal imaging of intracellular Ca2+ concentrations. Therefore, 6-NV-IP3 can be used for efficient, sub-cellular photorelease of IP3, not only in cultured cells but also, potentially, in vivo. It is in the latter situation that two-photon photolysis should reveal its true forte.
AB - The synthesis of a photolabile derivative of inositol-1,4,5-trisphosphate (IP3) is described. This new caged second messenger (6-ortho-nitroveratryl)-IP3 (6-NV-IP3) has an extinction coefficient of 5000M-1 cm-1 at 350 nm, and a quantum yield of photolysis of 0.12. Therefore, 6-NV-IP3 is photolyzed with UV light about three times more efficiently than the widely used P 4(5)-1-(2-nitrophenyl)ethyl-caged IP3 (NPE-IP 3). 6-NV-IP3 has a two-photon cross-section of about 0.035 GM at 730 nm. This absorbance is sufficiently large for effective two-photon excitation in living cells at modest power levels. Using near-IR light (5 mW, 710 nm, 80 MHz, pulse-width 70 fs), we produced focal bursts of IP3 in HeLa cells, as revealed by laser-scanning confocal imaging of intracellular Ca2+ concentrations. Therefore, 6-NV-IP3 can be used for efficient, sub-cellular photorelease of IP3, not only in cultured cells but also, potentially, in vivo. It is in the latter situation that two-photon photolysis should reveal its true forte.
KW - Caged compounds
KW - Calcium
KW - Photolysis
KW - Signal transduction
KW - Two-photon uncaging
UR - http://www.scopus.com/inward/record.url?scp=30444440460&partnerID=8YFLogxK
U2 - 10.1002/cbic.200500345
DO - 10.1002/cbic.200500345
M3 - Article
C2 - 16292788
AN - SCOPUS:30444440460
SN - 1439-4227
VL - 7
SP - 174
EP - 180
JO - ChemBioChem
JF - ChemBioChem
IS - 1
ER -