SUNBOR HOMEに戻る

公益財団法人 サントリー生命科学財団




生体機能システムや活性物質の同定・構造解析を通じて生命現象を理解し人類共通の安全と安心に貢献します

構成員

研究員野村 薫
研究員原田英里砂
 研究員 森 祥子

※構成員名をクリックでプロフィール(PDF)をご覧いただけます。

研究テーマ

1. 動的構造解析によるタンパク質の機能発現メカニズムの解明

生体機能の担い手であるタンパク質は、揺れ動くことによって機能します。例えば、酵素では、基質結合部位の構造を変化させながら酵素反応を進行させます。また、天然変性タンパク質に至っては、遊離状態ではランダムコイルであるにもかかわらず、選択的に標的分子と結合し、さらに特定の立体構造に折れ畳まれます。このようなタンパク質の揺らぎは、現在の構造生物学分野では当たり前として認識されるようになりましたが、従来の構造生物学では、主に静止した構造を研究しているため実際に機能を発現する際のタンパク質の実態とのギャップがあります。私たちはNMRを用いてタンパク質の揺らぎを解析し、揺らぎの面からタンパク質の機能を真に理解することを目指しています。

図1 図2

NMRでは原子分解能で、しかも広い時間領域(ナノ秒~時間)の揺らぎを解析できるため最もパワフルな揺らぎの解析法と言えます。緩和分散法と呼ばれる測定法を用いると、分子認識や構造変化の中間状態に存在する極めて低い存在比の状態(1%ほど)の構造情報まで取得できるようになってきました。私たちは、この緩和分散法などを用いて、これまでにヘム分解酵素のヘム認識機構を解析し、その結果、基質結合部位から離れた領域の揺らぎが活性に深く関わっていることを明らかにしました(図1)。

ヘム分解酵素以外にもiPS細胞を作製するのに用いられる転写因子Oct3/4とSox2のDNA認識機構(図2)の解明などにも取り組んでいます。DNA結合タンパク質では古くからDNA上でスライド(またはホップ)することによって、特異的な結合サイトを効率的に探索していることが知られています。Oct3/4とSox2の両者ともにαヘリックス構造と天然変性領域が存在しますが、私たちは、スライド状態から最終結合状態へ転移する過程に、この2つの領域の揺らぎがどのように関わるのかと言うことに焦点を絞って研究を進めています。

また揺らぎの解析は、方法論が十分に整備されているとは言えませんので、私たちは新規方法論の開発にも取り組んでいます。これまでに、緩和分散を1/10の時間で測定できる方法論の開発に成功しました。この方法論は不安定なタンパク質や濃度の薄いタンパク質の解析に広く有効ですので、更なる適用範囲の拡大を目指して、方法論の改良に取り組んでいます。

[ PAGE TOP ]

研究成果

【原著論文】

<2014>
1. Yanaka, S., Ueno, T., Shi, Y., Qi, J., Gao, G.F., Tsumoto, K., Sugase, K. (2014)
“Peptide-dependent Conformational Fluctuation Determines the Stability of the Human Leukocyte Antigen Class I Complex.” J. Biol. Chem. in press. [PubMed]
2. Walinda, E., Morimoto, D., Sugase, K., Konuma, T., Tochio, H., Shirakawa, M. (2014)
“Solution Structure of the Ubiquitin-associated (UBA) Domain of Human Autophagy Receptor NBR1 and its Interaction with Ubiquitin and Polyubiquitin.” J. Biol. Chem. 289, 13890-13902. [PubMed]
3. Nomura, K., Harada, E., Sugase, K., Shimamoto, K. (2014)
“Solid-state NMR spectra of lipid-anchored proteins under magic angle spinning.” J. Phys. Chem. B 118, 2405-2413. [PubMed]
4. Furukawa, A., Sugase, K., Morishita, R., Nagata, T., Kodaki, T., Takaori-Kondo, A., Ryo, A., Katahira, M. (2014)
“Quantitative analysis of location- and sequence-dependent deamination by APOBEC3G using real-time NMR spectroscopy.” Angew. Chem. Int. Ed. Engl. 53, 2349-2352. [PubMed]
<2013>
5. Sugase, K., Konuma, T., Lansing, J.C., Wright, P.E. (2013)
“Fast and accurate fitting of relaxation dispersion data using the flexible software package GLOVE.” J. Biomol. NMR 56, 275-283. [PubMed]
6. Kitazawa, S., Kameda, T., Yagi-Utsumi, M., Sugase, K., Baxter, N.J., Kato, K., Williamson, M.P., Kitahara, R. (2013)
“Solution structure of the Q41N variant of ubiquitin as a model for the alternatively folded N2 state of ubiquitin.” Biochemistry 52, 1874-1885. [PubMed]
7. Fukami, H., Yano, Y., Iwashita, T. (2013)
“Isolation of a reduced form of cyanidin 3-O-β-D-glucoside from immature black soybean (Glycine max (L.) Merr.) and its reducing properties.” J. Oleo. Sci. 62, 623-629. [PubMed]
<2012>
8. Yanagi, K., Sakurai, K., Yoshimura, Y., Konuma, T., Lee, Y.H., Sugase, K., Ikegami T., Naiki, H., Goto, Y. (2012)
“The monomer-seed interaction mechanism in the formation of the β2-microglobulin amyloid fibril clarified by solution NMR techniques.” J. Mol. Biol. 422, 390-402. [PubMed]
9. Nishiyama, K., Maeda, M., Yanagisawa, K., Nagase, R., Komura, H., Iwashita, T., Yamagaki, T., Kusumoto, S., Tokuda, H., Shimamoto, K. (2012)
“MPIase is a glycolipozyme essential for membrane protein integration.” Nat. Commun. 3, 1260. [PubMed]
10. Kajikawa, T., Okumura, S., Iwashita, T., Kosumi, D., Hashimoto, H., Katsumura, S. (2012)
“Stereocontrolled total synthesis of fucoxanthin and its polyene chain-modified derivative.” Org. Lett. 14, 808-811. [PubMed]
<2011>
11. Matsuki, Y., Konuma, T. Fujiwara, T., Sugase, K. (2011)
“Boosting protein dynamics studies using quantitative nonuniform sampling NMR spectroscopy.” J. Phys. Chem. B. 115, 13740-13745. [PubMed]
12. Sugase, K. (2011)
“Elucidating slow binding kinetics of a protein without observable bound resonances by longitudinal relaxation NMR spectroscopy.” J. Biomol. NMR 50, 219-227. [PubMed]
<2010>
13. Ono, E., Ruike, M., Iwashita, T., Nomoto K., Fukui, Y. (2010)
“Co-pigmentation and flavonoid glycosyltransferases in blue Veronica persica flowers.” Phytochem. 71, 726-735. [PubMed]
14. Ando, H., Ueoka, R., Okada, S., Fujita, T., Iwashita, T., Imai, T., Yokoyama, T., Matsumoto, Y., van Soest, R.W., Matsunaga, S. (2010)
“Penasins A-E, long-chain cytotoxic sphingoid bases, from a marine sponge Penares sp.” J. Nat. Prod. 73, 1947-1950. [PubMed]
15. Igarashi, Y., Kim, Y., In Y., Ishida, T., Kan Y., Fujita, T., Iwashita, T., Tabata, H., Onaka, H., Furumai, T. (2010)
“Alchivemycin A, a bioactive polycyclic polyketide with an unprecedented skeleton from Streptomyces sp.” Org. Lett. 12, 3402-3405. [PubMed]
16. Kajikawa, T., Aoki, K., Iwashita, T., Niedzwiedzki, D.M., Frank, H.A., Katsumura, S. (2010)
“Syntheses of ylidenbutenolide-modified derivatives of peridinin and their stereochemical and spectral characteristics.” Org. Biomol. Chem. 8, 2513-2516. [PubMed]
17. Namba ,K., Kobayashi, K., Murata, Y., Hirakawa, H., Yamagaki, T., Iwashita, T., Nishizawa, M., Kusumoto, S., Tanino, K. (2010)
“Mugineic acid derivatives as molecular probes for the mechanistic elucidation of iron acquisition in barley.” Angew. Chem. Int. Ed. Engl. 49, 9956-9959. [PubMed]
<2009>
18. Yamaji, N., Little, M.J., Nishio, H., Billen, B., Villegas, E., Nishiuchi, Y., Tytgat, J., Nicholson, G.M., Corzo, G. (2009)
“Synthesis, solution structure, and phylum selectivity of a spider delta-toxin that slows inactivation of specific voltage-gated sodium channel subtypes.” J. Biol. Chem. 284, 24568-24582. [PubMed]
19. Washida, K., Yamagaki, T., Iwashita, T., Nomoto, K. (2009)
“Two new galloylated monoterpene glycosides, 4-O-galloylalbiflorin and 4'-O-galloylpaeoniflorin, from the roots of Paeonia lactiflora (Paeoniae radix) grown and processed in Nara prefecture, Japan.” Chem. Pharm. Bull. 57, 1150-1152. [PubMed]
20. Kajikawa, T., Hasegawa, S., Iwashita, T., Kusumoto, T., Hashimoto, H., Niedzwiedzki, D.M., Frank, H.A., Katsumura, S. (2009)
“Syntheses of C33-, C35-, and C39-peridinin and their spectral characteristics.” Org. Lett. 11, 5006-5009. [PubMed]
21. Washida, K., Itoh, Y., Iwashita, T., Nomoto, K.
“Androgen modulators from the roots of Paeonia lactiflora (Paeoniae radix) grown and processed in Nara prefecture, Japan.” Chem. Pharm. Bull. 57, 971-974. [PubMed]
22. Kajikawa, T., Aoki, K., Singh, R.S., Iwashita, T., Kusumoto, T., Frank, H.A., Hashimoto, H., Katsumura, S. (2009)
“Syntheses of allene-modified derivatives of peridinin toward elucidation of the effective role of the allene function in high energy transfer efficiencies in photosynthesis.” Org. Biomol. Chem. 7, 3723-3733. [PubMed]
<2008>
23. Sugase, K., Landes, M.A., Wright, P.E., Martinez-Yamout, M. (2008)
“Overexpression of post-translationally modified peptides in Escherichia coli by co-expression with modifying enzymes.” Protein Expr. Purif. 57, 108-115. [PubMed]
24. Usuki, H., Nitoda, T., Ichikawa, M., Yamaji, N., Iwashita, T., Komura, H., Kanzaki, H. (2008)
“TMG-chitotriomycin, an enzyme inhibitor specific for insect and fungal beta-N-acetylglucosaminidases, produced by actinomycete Streptomyces anulatus NBRC 13369.” J. Am. Chem. Soc. 130, 4146-4152. [PubMed]
25. Kim, D., Miyazaki, Y., Nakashima, T., Iwashita, T., Fujita, T., Yamaguchi, K., Choi, K. S., Oda, T. (2008)
“Cytotoxic action mode of a novel porphyrin derivative isolated from harmful red tide dinoflagellate Heterocapsa circularisquama.” J. Biochem. Mol. Toxicol. 22, 158-165. [PubMed]
26. Ueno, D., Iwashita, T., Zhao, F.J., Ma, J.F. (2008)
“Characterization of Cd translocation and identification of the Cd form in xylem sap of the Cd-hyperaccumulator Arabidopsis halleri.” Plant Cell Physiol. 49, 540-548. [PubMed]
27. Nakashima, T., Iwashita, T., Fujita, T., Sato, E., Niwano, Y., Kohno, M., Kuwahara, S., Harada, N., Takeshita, S., Oda, T. (2008)
“A prodigiosin analogue inactivates NADPH oxidase in macrophage cells by inhibiting assembly of p47phox and Rac.” J. Biochem. 143, 107-115. [PubMed]
28. Ueoka R., Fujita T., Iwashita T., van Soest R.W., Matsunaga S. (2008)
“Inconspicamide, new N-acylated serinol from the marine sponge Stelletta inconspicua.” Biosci. Biotechnol. Biochem. 72, 3055-3058. [PubMed]
<2007>
29. Sugase, K., Lansing, J.C., Dyson, H.J., Wright, P.E. (2007)
“Tailoring relaxation dispersion experiments for fast-associating protein complexes.” J. Am. Chem. Soc. 129, 13406-13407. [PubMed]
30. Yamaji, N., Sugase, K., Nakajima, T., Miki, T., Wakamori, M., Mori, Y., Iwashita, T. (2007)
“Solution structure of agelenin, an insecticidal peptide isolated from the spider Agelena opulenta, and its structural similarities to insect-specific calcium channel inhibitors.” FEBS Lett. 581, 3789-3794. [PubMed]
31. Sugase, K., Dyson, H.J., Wright, P.E. (2007)
“Mechanism of coupled folding and binding of an intrinsically disordered protein.” Nature 447, 1021-1025. [PubMed]
32. Okamoto, C., Nakao, Y., Fujita, T., Iwashita, T., van Soest, R.W., Fusetani, N., Matsunaga, S. (2007)
“Cytotoxic C47-polyacetylene carboxylic acids from a marine sponge Pertrosia sp.” J. Nat. Prod. 70, 1816-1819. [PubMed]
33. Ueno, D., Rombolà, A.D., Iwashita, T., Nomoto, K., Ma, J.F. (2007)
“Identification of two novel phytosiderophores secreted by perennial grasses.” New Phytol. 174, 304-310. [PubMed]
34. Yamaji, N., Yokoo, Y., Iwashita, T., Nemoto, A., Koike, M., Suwa, Y., Wakimoto, T., Tsuji, K., Nukaya, H. (2007)
“Structural determination of two active compounds that bind to the muscarinic M3 receptor in beer.” Alcohol Clin. Exp. Res. 31, S9-S14. [PubMed]
35. Kobayashi, H., Ohashi, J., Fujita, T., Iwashita, T., Nakao, Y., Matsunaga, S., Fusetani, N. (2007)
"Complete Structure elucidation of shishididemniols, complex lipids with tyramine-derived tether and two serinol units, from a marine tunicate of the family Didemnidae." J. Org. Chem. 72, 1218-1225. [PubMed]
<2006>
36. Dmitrenok, A., Iwashita, T., Nakajima, T., Sakamoto, B., Namikoshi, M., and Nagai, H. (2006)
“New cyclic depsipeptides from the green alga Bryopsis species; application of a carboxypeptidase hydrolysis reaction to the structure determination.” Tetrahedron 62, 1301-1308. [publisher]
37. Fukui, Y., Nomoto, K., Iwashita, T., Masuda, K., Tanaka, Y., and Kusumi, T. (2006)
“Two novel blue pigments with ellagitannin moiety, rosacyanins A1 and A2, isolated from the petals of Rosa hybrid.” Tetrahedron 62, 9661-9670. [publisher]
38. Hao, J., Matsuura, F., Kishi, Y., Kita, M., Uemura, D., Asai, N., Iwashita, T. (2006)
“Stereochemistry of pteriatoxins A, B, and C.” J. Am. Chem. Soc. 128, 7742-7743. [PubMed]
39. Morsy, N., Houdai, T., Matsuoka, S., Matsumori, N., Adachi, S., Oishi, T., Murata, M., Iwashita, T., Fujita, T. (2006)
“Structures of new amphidinols with truncated polyhydroxyl chain and their membrane-permeabilizing activities.” Bioorg. Med. Chem. 14, 6548-6554. [PubMed]
40. Miyazaki, Y., Iwashita, T., Yamaguchi, K., Oda, T., Matsuyama, Y., Honjo, T. (2006)
“Isolation and characterization of photoactive haemolytic toxin from Heterocapsa circularisquama.” Afr. J. Mar. Sci. 28, 427-429. [publisher]
<2005以前>
41. Nomura, K., Ferrat, G., Nakajima, T., Darbon, H., Iwashita, T., Corzo, G. (2005)
“Induction of morphological changes in model lipid membranes and the mechanism of membrane disruption by a large scorpion-derived pore-forming peptide.” Biophys. J., 89, 4067-4080. [PubMed]
42. Miyazaki, Y., Nakashima, T., Iwashita, T., Fujita, T., Yamaguchi, K., Oda, T. (2005)
“Purification and characterization of photosensitizing hemolytic toxin from harmful red tide phytoplankton, Heterocapsa circularisquama.” Aquat Toxicol. 73, 382-393.[PubMed]
43. Ueno, D., Ma, J.F., Iwashita, T., Zhao F.J., McGrath S.P. (2005)
“Identification of the form of Cd in the leaves of a superior Cd-accumulating ecotype of Thlaspi caerulescens using 113Cd-NMR.” Planta. 221, 928-936. [PubMed]
44. Ma, J.F., Mitani, N., Nagao, S., Konishi, S., Tamai, K., Iwashita, T., Yano, M. (2004)
“Characterization of the silicon uptake system and molecular mapping of the silicon transporter gene in rice.” Plant Physiol. 136, 3284-3289. [PubMed]
45. Nomura, K., Corzo, G., Nakajima, T., Iwashita, T. (2004)
“Orientation and pore-forming mechanism of a scorpion pore-forming peptide bound to magnetically oriented lipid bilayers.” Biophys J. 87, 2497-2507. [PubMed]
46. Yamaji, N., Dai, L., Sugase, K., Andriantsiferana, M., Nakajima, T., Iwashita, T. (2004)
“Solution structure of IsTX. A male scorpion toxin from Opisthacanthus madagascariensis (Ischnuridae).” Eur. J. Biochem. 271, 3855-3864. [PubMed]
47. Sugase, K., Horikawa, M., Sugiyama M, Ishiguro, M. (2004)
“Restriction of a peptide turn conformation and conformational analysis of guanidino group using arginine-proline fused amino acids: application to mini atrial natriuretic peptide on binding to the receptor.” J. Med. Chem. 47, 489-492 [PubMed]
48. Shen, R., Iwashita, T., Ma, J.F. (2004)
“Form of Al changes with Al concentration in leaves of buckwheat.” J. Exp. Bot. 55, 131-136. [PubMed]
49. Yamaji, N., Horikawa, M., Corzo, G., Naoki, H., Haupt, J., Nakajima, T., Iwashita, T. (2004)
“Structure and enantioselective synthesis of polyamine toxin MG30 from the venom of the spider Macrothele gigas.” Tetrahedron Lett. 45, 5371-5373. [publisher]
50. Fukui, Y., Tanaka, Y., Kusumi, T., Iwashita, T., Nomoto, K. (2003)
“A rationale for the shift in colour towards blue in transgenic carnation flowers expressing the flavonoid 3’,5’-hydroxylase gene.” Phytochemistry 63, 15-23. [PubMed]
51. Kato-Toma, Y., Iwashita, T., Masuda K., Oyama, Y., Ishiguro, M. (2003)
“pKa measurements from nuclear magnetic resonance of tyrosine-150 in class C -lactamase.” Biochem. J. 371, 175-181. [PubMed]
52. Fukui, Y., Kusumi, T., Masuda, K., Iwashita, T., Nomoto, K. (2002)
“Structure of rosacyanin B, a novel pigment from the petals of Rosa hybrida.” Tetrahedron Lett. 43, 2637-2639. [publisher]
53. Sugase, K., Oyama, Y., Kitano, K., Akutsu, H., Ishiguro, M. (2002)
“Structure-activity relationships for mini atrial natriuretic peptide by proline-scanning mutagenesis and shortening of peptide backbone.” Bioorg. Med. Chem. Lett. 12, 1245-1247. [PubMed]
54. Sugase, K., Oyama, Y., Kitano, K., Iwashita, T., Fujiwara, T., Akutsu, H., Ishiguro, M. (2002)
“Designing analogues of mini atrial natriuretic peptide based on structural analysis by NMR and restrained molecular dynamics.” J. Med. Chem. 45, 881-887. [PubMed]
55. Kan, Y., Uemura, D., Hirata, Y., Ishiguro, M., Iwashita, T. (2001)
“Complete NMR signal assignment of palytoxin and N-acetylpalytoxin.” Tetrahedron Lett. 42, 3197-3202. [publisher]
56. Okuno, Y., Iwashita, T., Sugiura, Y. (2000)
“Structural basis for reaction mechanism and drug delivery system of chromoprotein antitumor antibiotic C-1027.” J. Am. Chem. Soc. 122, 6848-6854. [publisher]
57. Li, Y.Q., Sugase, K., Ishiguro, M. (1999)
“Synthesis of the cyclic heptapeptide Substance P antagonist, dihydro-WIN67689 and determination of the stereochemistry of the modified tyrosine moiety.” Tetrahedron Lett. 40, 9097-9100. [publisher]

[ PAGE TOP ]

【総説・著書】

<2013>
58. 菅瀬 謙治 (2013)
「5.2 緩和分散法」揺らぎ・ダイナミクスと生体機能 物理化学的視点から見た生体分子(寺嶋正秀 編), 79-90 [publisher]
59. 菅瀬 謙治 (2013)
「実践relaxation dispersion法」 NMR 3, 82-85. [publisher]
<2012>
60. 合田 麻美, 堀川 学, 岩下 孝, 澤田 元充, 塚嵜 大輔, 橘 津義, 菅根 克己, 原田 雅己 (2012)
「赤色102号中に微量存在するアゾ化合物の構造解析」食衛誌 54, 188-197. [publisher]
61. 岩下 孝 (2012)
「特論NMR立体化学」講談社サイエンティフィク [Amazon]
<2009>
62. Wright, P.E., Felitsky, D. J., Sugase, K., Dyson, H.J. (2009)
“Mapping Protein Folding Landscapes by NMR Relaxation” Water and Biomolecules, Springer-Verlag, 1-12. [publisher]
<2008>
63. 菅瀬 謙治 (2008)
「Relaxation Dispersion法の生化学への応用」生化学 80, 754-758. [publisher]
<2007>
64. 菅瀬 謙治 (2007)
「天然変性状態と遭遇複合体」タンパク質核酸酵素 52, 945-951. [PubMed]
65. 菅瀬 謙治 (2007)
「レスベラトロールがメタボリックシンドロームを改善する?」ファルマシア 43, 721-722. [publisher]
<取材等>
1. 日経産業新聞
平成19年5月24日11面

[ PAGE TOP ]