Department of Biomolecular Science

Toshiaki Furuta, Organic and Bioorganic Chemistry,

Controlling cellular chemistry using caged compounds

Our research program focuses on developing small molecules that can manipulate biological processes in live cells and tissue samples. Live cell imaging would tell us how cells respond to various external stimuli. In such experiments, artificial stimulation is needed to add environmental perturbation to cells. The stimulation must be applied in a way so as to mimic physiological conditions, otherwise it is ambiguous whether observed responses are physiologically important. We have been working on developing novel methods which allow us to control cellular chemistry without causing any damages to live cells and tissues. Photo-stimulation of caged compounds has potential usefulness in such experiments. Caged compounds are synthetic molecules whose biological activities are temporarily masked by the covalent attachment of photochemically removable protecting groups. The original activity of the molecule will be restored once photo-irradiation has been applied. Therefore, localized stimulation of a particular biological process is possible within defined time windows. Bhc, 6-bromo-7-hydroxycoumarin-4-ylmethyl, group is a newly developed photoremovable protecting group, which has practically high one and two-photon uncaging efficiencies and has been used to make caged compounds of neurotransmitters, second messengers, mRNAs and so forth.

Recent Publications

T. Watanabe, T. Hoshida, J. Sakyo, M. Kishi, S. Tanabe, J. Matsuura, S. Akiyama, M. Nakata, Y. Tanabe, A. Suzuki, S. Watanabe, T. Furuta, Synthesis of nucleobase-caged peptide nucleic acids having improved photochemical properties, Org. Biomol. Chem. 12, 5089-5093 (2014).

 H. Takano, T. Narumi, N. Ohashi, A. Suzuki, T. Furuta, W. Nomura, H. Tamamura, Development of the 8-aza-3-bromo-7-hydroxycoumarin-4-ylmethyl group as a new entry of photolabile protecting groups, Tetrahedron, 70, 4400-4404 (2014).

 T. Narumi, H. Takano, N. Ohashi, A. Suzuki, T. Furuta, H. Tamamura, Isostere-Based Design of 8-Azacoumarin-type Photolabile Protecting Groups: A Hydrophilicity Increasing Strategy for Coumarin-4-ylmethyls, Org. Lett. 16, 1184-1187 (2014).

 A. Teraoka, K. Murakoshi, K. Fukamauchi, A. Z. Suzuki, S. Watanabe, T. Furuta, Preparation and affinity-based purification of caged linear DNA for light-controlled gene expression in mammalian cells, Chem. Commun. 50, 664-666 (2014).

T. Furuta, K. Manabe, A. Teraoka, K. Murakoshi, A. Ohtsubo, A. Suzuki, Design, synthesis and photochemistry of modular caging groups for caged nucleotides, Org. Lett. 14, 6182-6185 (2012).

 T. Furuta, Designing caged compounds for spatiotemporal control of cellular chemistry, Journal of Synthetic Organic Chemistry, Japan, 69, 1164-1169 (2012).

 W. Nomura, T. Narumi, N. Ohashi, Y. Serizawa, N. E. Lewin, P. M. Blumberg, T. Furuta, H. Tamamura, Synthetic Caged DAG-lactones for Photochemically Controlled Activation of Protein Kinase C, ChemBioChem, 12, 535–539 (2011).

 S. Mizukami, M. Hosoda, T. Satake, S. Okada, Y. Hori, T. Furuta and K. Kikuchi, Photocontrolled Compound Release System Using Caged Antimicrobial Peptide, J. Am. Chem. Soc. 132, 9524-9525 (2010).

 A. Shigenaga, J. Yamamoto, Y. Sumikawa, T. Furuta, A. Otaka, Development and photo-sensitive peptide bond cleavage reaction of two-photon near infrared excitation-responsive peptide, Tetrahedron Lett. 51, 2868-2871 (2010).

 S. Yamaguchi, Y. Chen, S. Nakajima, T. Furuta, T. Nagamune, Light-activated gene expression from site-specific caged DNA with a biotinylated photolabile protection group, Chem. Commun. 46, 2244--2246, (2010).

 K. Hishikawa, H. Nakagawa, T. Furuta, K. Fukuhara, H. Tsumoto, T. Suzuki, N. Miyata, Multiple bond-conjugated photoinduced nitric oxide releaser working with two-photon excitation. Bioorg. Med. Chem. Lett. 20, 302-305 (2010).

 H. Tsutsui, H. Shimizu, H. Mizuno, N. Nukina, T. Furuta and A. Miyawaki, The E1 mechanism in photo-induced β-elimination reactions for green-to-red conversion of fluorescent proteins, Chem. & Biol. . 16, 1140-1147 (2009).

 K. Hishikawa, H. Nakagawa, T. Furuta, K. Fukuhara, H. Tsumoto, T. Suzuki, N. Miyata, Photoinduced nitric oxide release from a hindered nitrobenzene derivative by two-photon excitation., J. Am. Chem. Soc. 131, 7488-7489 (2009).

 E. J. Quann, E. Merino, T. Furuta, and M Huse, Localized diacylglycerol drives the polarization of the microtubule-organizing center in T cells, Nature Immun. 10, 627-635 (2009).

 K. Katayama, S. Tsukiji, T. Furuta, and T. Nagamune, A bromocoumarin-based linker for synthesis of photocleavable peptidoconjugates with high photosensitivity. Chem Commun, 5399-5401 (2008).

T. Kawakami, H. Cheng, S. Hashiro, Y. Nomura, S. Tsukiji, T. Furuta, T. Nagamune, A Caged Phosphopeptide-Based Approach for Photochemical Activation of Kinases in Living Cells, ChemBioChem, 9, 1583-1586 (2008)

C. Orang, A. Specht, D. Puliti, E. Sakr, T. Furuta, B. Winsor, M. Goeldner, Synthesis and Photochemical Properties of a Light-activated Fluorophore to label His-tagged Proteins, Chem. Commun. 1217-1219 (2008)

T. Furuta, S. Tanabe, T. Watanabe, J. Sakyo, C. Matsuba, Phototriggers for Nucleobases with Improved Photochemicasl Properties Org. Lett. 9, 4717-4720 (2007)

I. Hayashi, H. Mizuno, K. I. Tong, T. Furuta, F. Tanaka, M. Yoshimura, A. Miyawaki, M. Ikura, Crystallographic Evidence for Water-assisted Photo-induced Peptide Cleavage in the Stony Coral Fluorescent Protein Kaede, J. Mol. Biol., 372, 918-926 (2007).

A. Specht, J. S. Thomann, K. Alarcon, W. Wittayanan, D. Ogden, T. Furuta, Y. Kurakawa, M. Goeldner, New Photoremovable Protecting Groups for Carboxylic Acids with High Photolytic Efficiencies at Near-UV Irradiation. Application to the Photocontrolled Release of L-Glutamate. ChemBioChem. 7, 1690-1695 (2006).

H. Ando, M. Kobayashi, T. Tsubokawa, K. Uyemura, T. Furuta, H. Okamoto, Lhx2 mediates the activity of Six3 in zebrafish forebrain growth, Dev. Biol. 287, 456-468 (2005).

C. D. Wood, T. Nishigaki, T. Furuta, S. A. Baba, A. Darszon, Real-time analysis of the role of Ca2+ in flagellar movement and motility in single sea urchin sperm, J. Cell. Biol. 169, 725-731 (2005).

T. Furuta, K. Noguchi, Controlling cellular systems using Bhc-caged compounds, Trends. Anal. Chem. 23, 501-509 (2004).

T. Furuta, H. Takeuchi, M. Isozaki, Y. Takahashi, M. Sugimoto, M .Kanehara, T. Watanabe, K. Noguchi, T. M. Dore, T. Kurahashi, M. Iwamura, R. Y. Tsien, Bhc-cNMPs as either water-soluble or membrane-permeant photo-releasable cyclic nucleotides for both one and two-photon excitation, ChemBioChem, 5, 1119-1128 (2004).

T. Nishigaki, C. D. Wood, Y. Tatsu, N. Yumoto, T. Furuta, D. Ellias, K. Shiba, S. A. Baba, A. Darszon, A sea urchin jelly peptide induces a cGMP-mediated decrease in sperm intracellular Ca2+ before its increase, Dev. Biol. 272, 376-388 (2004).

H. Mizuno, T. K. Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, A. Miyawaki, Photo-induced peptide cleavage for the green-to-red conversion of a fluorescent protein, Mol. Cell 12, 1051-1058 (2003)

A. Z. Suzuki, T. Watanabe, M. Kawamoto, K. Nishiyama, H. Yamashita, M. Iwamura and T. Furuta, Coumarin-4-ylmethoxycarbonyls as Phototriggers for Alcohols and Phenols, Org. Lett., 5, 4867-4870 (2003).

Y. Hirayama, M. Iwamura and T. Furuta, Design, Synthesis and Photochemical Properties of Caged Bile Acids, Bioorg. Med. Chem. Lett., 13, 905-908 (2003).

H. Ando, T. Furuta, R. Y. Tsien and H. Okamoto, Photo-mediated gene activation using caged RNA/DNA in zebrafish embryos. Nature Genetics, 28, 317-325 (2001).

 T. Furuta, Y. Hirayama and M. Iwamura, Anthraquinone-2-ylmethoxycarbonyl (Aqmoc): a new photochemically removable protecting group for alcohols. Org. Lett. 3, 1809-1812 (2001).

T. Furuta, S. S.-H. Wang, J. L. Dantzker, T. M. Dore, W. J. Bybee, E. M. Callaway, W. Denk and R. Y. Tsien Brominated 7-Hydroxycoumarin-4-ylmethyls: Photolabile Protecting Groups with Biologically Useful Cross-sections for Two Photon Photolysis, Proc. Natl. Acad. Sci. USA, 96, 1193-1200 (1999).