Spatiotemporal laser inactivation of inositol 1,4,5-trisphosphate receptors using synthetic small-molecule probes

Takanari Inoue, Kazuya Kikuchi, Kenzo Hirose, Masamitsu Iino, Tetsuo Nagano

Research output: Contribution to journalArticlepeer-review

Abstract

A malachite green-conjugated inositol 1,4,5-trisphosphate (MGIP3) induces specific inactivation of IP3 receptor (IP3R) in tissue samples upon laser irradiation. To verify potential usefulness of the method for studies of cellular Ca2+ signaling, we conducted laser inactivation at the single-cell level and show that IP3R was inactivated with extremely high spatiotemporal resolution. In the presence of MGIP3, the Ca2+ release function of IP3R in single B lymphoma cells decayed exponentially with increasing duration of laser irradiation with a time constant of 3.4 s. Moreover, by confining laser irradiation to a spatially distinct region of differentiated PC12 cells, subcellular inactivation of IP3R was attained, as revealed by a loss of local Ca2+ signal. Such real-time inactivation of IP3R only within a subcellular region may provide a powerful method for investigating spatiotemporal dynamics of Ca2+ signaling.

Original languageEnglish (US)
Pages (from-to)503-509
Number of pages7
JournalChemistry and Biology
Volume10
Issue number6
DOIs
StatePublished - Jun 1 2003
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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