Synthesis and two-photon photolysis of 6-(ortho-nitroveratryl)-caged IP3 in living cells

Srinivas Kantevari, Caroline J. Hoang, Jakob Ogrodnik, Marcel Egger, Ernst Niggli, Graham C.R. Ellis-Davies

Research output: Contribution to journalArticlepeer-review

48 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)174-180
Number of pages7
JournalChemBioChem
Volume7
Issue number1
DOIs
StatePublished - Jan 2006
Externally publishedYes

Keywords

  • Caged compounds
  • Calcium
  • Photolysis
  • Signal transduction
  • Two-photon uncaging

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