A novel fluorescent cell membrane-permeable caged cyclic ADP-ribose analogue

J Biol Chem. 2012 Jul 13;287(29):24774-83. doi: 10.1074/jbc.M111.329854. Epub 2012 Jun 1.


Cyclic adenosine diphosphate ribose is an endogenous Ca(2+) mobilizer involved in diverse cellular processes. A cell membrane-permeable cyclic adenosine diphosphate ribose analogue, cyclic inosine diphosphoribose ether (cIDPRE), can induce Ca(2+) increase in intact human Jurkat T-lymphocytes. Here we synthesized a coumarin-caged analogue of cIDPRE (Co-i-cIDPRE), aiming to have a precisely temporal and spatial control of bioactive cIDPRE release inside the cell using UV uncaging. We showed that Co-i-cIDPRE accumulated inside Jurkat cells quickly and efficiently. Uncaging of Co-i-cIDPRE evoked Ca(2+) release from endoplasmic reticulum, with concomitant Ca(2+) influx in Jurkat cells. Ca(2+) release evoked by uncaged Co-i-cIDPRE was blocked by knockdown of ryanodine receptors (RyRs) 2 and 3 in Jurkat cells. The associated Ca(2+) influx, on the other hand, was abolished by double knockdown of Stim1 and TRPM2 in Jurkat cells. Furthermore, Ca(2+) release or influx evoked by uncaged Co-i-cIDPRE was recapitulated in HEK293 cells that overexpress RyRs or TRPM2, respectively, but not in wild-type cells lacking these channels. In summary, our results indicate that uncaging of Co-i-cIDPRE incites Ca(2+) release from endoplasmic reticulum via RyRs and triggers Ca(2+) influx via TRPM2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkenes / metabolism
  • Blotting, Western
  • Calcium
  • Cell Line
  • Cell Membrane / metabolism*
  • Coumarins / metabolism
  • Cyclic ADP-Ribose / analogs & derivatives*
  • Cyclic ADP-Ribose / metabolism
  • Fluorescence
  • HEK293 Cells
  • Humans
  • Jurkat Cells
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Real-Time Polymerase Chain Reaction
  • Ryanodine Receptor Calcium Release Channel / genetics
  • Ryanodine Receptor Calcium Release Channel / metabolism
  • Stromal Interaction Molecule 1
  • TRPM Cation Channels / genetics
  • TRPM Cation Channels / metabolism


  • Alkenes
  • Coumarins
  • Membrane Proteins
  • Neoplasm Proteins
  • Ryanodine Receptor Calcium Release Channel
  • STIM1 protein, human
  • Stromal Interaction Molecule 1
  • TRPM Cation Channels
  • TRPM2 protein, human
  • caged cyclic ADP-ribose
  • Cyclic ADP-Ribose
  • coumarin
  • propylene
  • Calcium