Bhc-cNMPs as either water-soluble or membrane-permeant photoreleasable cyclic nucleotides for both one- and two-photon excitation

Toshiaki Furuta, Hiroko Takeuchi, Masahiro Isozaki, Yasuhiro Takahashi, Makoto Kanehara, Masazumi Sugimoto, Takayoshi Watanabe, Kousei Noguchi, Timothy M. Dore, Takashi Kurahashi, Michiko Iwamura, Roger Y. Tsien

Research output: Contribution to journalArticlepeer-review

Abstract

Cyclic nucleoside monophosphates (cNMPs) play key roles in many cellular regulatory processes, such as growth, differentiation, motility, and gene expression. Caged derivatives that can be activated by irradiation could be powerful tools for studying such diverse functions of intracellular second messengers, since the spatiotemporal dynamics of these molecules can be controlled by irradiation with appropriately focused light. Here we report the synthesis, photochemistry, and biological testing of 6-bromo-7-hydroxycoumarin- 4-ylmethyl esters of cNMP (Bhc-cNMP) and their acetyl derivatives (Bhc-cNMP/Ac) as new caged second messengers. Irradiation of Bhc-cNMPs quantitatively produced the parent cNMPs with one-photon uncaging efficiencies (Φε) of up to one order of magnitude better than those of 2-nitrophenethyl (NPE) cNMPs. In addition, two-photon induced photochemical release of cNMP from Bhc-cNMPs (7 and 8) can be observed with the two-photon uncaging action cross-sections (δu) of up to 2.28 GM (1 GM = 10-50 cm4 s photon-1), which is the largest value among those of the reported Bhc-caged compounds. The wave-length dependence of the δu values of 7 revealed that the peak wavelength was twice that of the one-photon absorption maximum. Bhc-cNMPs showed practically useful water solubility (nearly 500 μM), whereas 7-acetylated derivatives (Bhc-cNMPs/Ac) were expected to have a certain membrane permeability. Their advantages were demonstrated in two types of biological systems: the opening of cAMP-mediated transduction channels in newt olfactory receptor cells and cAMP-mediated motility responses in epidermal melanophores in scales from medaka fish. Both examples showed that Bhc and Bhc/Ac caged compounds have great potential for use in many cell biological applications.

Original languageEnglish (US)
Pages (from-to)1119-1128
Number of pages10
JournalChemBioChem
Volume5
Issue number8
DOIs
StatePublished - Aug 6 2004

Keywords

  • Cage compounds
  • Nucleotides
  • Protecting groups
  • Signal transduction
  • Two-photon excitation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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