Publication List of Toshio Iwasaki Group Homepage|Nippon Medical School|日本医科大学|岩崎 俊雄 グループ ホームページ

Toshio Iwasaki, Ph.D. (Principal Investigator) LinkIconGo to the ICC Project Page

Position: Group Leader and Lecturer
Address: Department of Biochemistry and Molecular Biology,
Nippon Medical School
1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, JAPAN
(Lab) Room 3D08, 1-25-16 Nezu, Bunkyo-ku, Tokyo 113-0031, JAPANLinkIcon

Fax: +81-(0)3-5685-3054
E-mail: tiwasaki(at)nms.ac.jp


Research interests: Biochemistry, Biophysics, Bioinorganic chemistry, and Biology

Brief description of the work: Structural, functional, and evolutionary biochemistry applied to simple and complex metalloproteins in hyperthermophiles and mammals

Original papers (refereed) [*corresponding author(s)] - where possible, include links for you to read them.

1. Iwasaki, T.,* Wakagi, T., Isogai, Y., Tanaka, K., Iizuka, T., and Oshima, T. (1994) Functional and evolutionary implications of a [3Fe-4S] cluster of the dicluster-type ferredoxin from the thermoacidophilic archaeon, Sulfolobus sp. strain 7. J. Biol. Chem. 269, 29444-29450. Pubmed

2. Iwasaki, T.,* Matsuura, K., and Oshima, T. (1995) Resolution of the aerobic respiratory system of the thermoacidophilic archaeon, Sulfolobus sp. strain 7: I. the archaeal terminal oxidase super-complex is a functional fusion of respiratory complexes III and IV with no c-type cytochromes. J. Biol. Chem. 270, 30881-30892. Pubmed

3. Iwasaki, T.,* Wakagi, T., Isogai, Y., Iizuka, T., and Oshima, T. (1995) Resolution of the aerobic respiratory system of the thermoacidophilic archaeon, Sulfolobus sp. strain 7: II. characterization of the archaeal terminal oxidase subcomplexes and implication for the intramolecular electron transfer. J. Biol. Chem. 270, 30893-30901. Pubmed

4. Iwasaki, T.,* Wakagi, T., and Oshima, T. (1995) Resolution of the aerobic respiratory system of the thermoacidophilic archaeon, Sulfolobus sp. strain 7: III. the archaeal novel respiratory complex II (succinate:caldariellaquinone oxidoreductase complex) inherently lacks heme group. J. Biol. Chem. 270, 30902-30908. Pubmed

5. Iwasaki, T.,* Wakagi, T., and Oshima, T. (1995) Ferredoxin-dependent redox system of a thermoacidophilic archaeon, Sulfolobus sp. strain 7: purification and characterization of a novel reduced ferredoxin-reoxidizing iron-sulfur flavoprotein. J. Biol. Chem. 270, 17878-17883. Pubmed

6. Iwasaki, T.,* Isogai, Y., Iizuka, T., and Oshima, T. (1995) Sulredoxin: a novel iron-sulfur protein of the thermoacidophilic archaeon Sulfolobus sp. strain 7 with a Rieske-type [2Fe-2S] cluster. J. Bacteriol. 177, 2576-2582. Pubmed

7. Iwasaki, T.,* Fujii, T., Wakagi, T., and Oshima, T. (1995) Alternative form of the dicluster-type ferredoxin from the thermoacidophilic archaeon, Sulfolobus sp. strain 7. Biochem. Biophys. Res. Commun. 206, 563-569. Pubmed

8. Iwasaki, T.,* Imai, T., Urushiyama, A., and Oshima, T. (1996) Redox-linked ionization of sulredoxin of Sulfolobus sp. strain 7, an archaeal Rieske-type [2Fe-2S] protein. J. Biol. Chem. 271, 27659-27663. Pubmed

9. Zhang, Q., Iwasaki, T., Wakagi, T.,* and Oshima, T. (1996) 2-Oxoacid:ferredoxin oxidoreductase from the thermoacidophilic archaeon, Sulfolobus sp. strain 7. J. Biochem. 120, 587-599. Pubmed

10. Iwasaki, T.,* and Oshima, T. (1996) Role of cytochrome b562 in the archaeal aerobic respiratory chain of Sulfolobus sp. strain 7. FEMS Microbiol. Lett. 144, 259-266. Pubmed

11. Suzuki, T.,* Inoki, Y., Yamagishi, A., Iwasaki, T., Wakagi, T., and Oshima, T. (1997) Molecular and phylogenetic characterization of isopropylmalate dehydrogenase of a thermoacidophilic archaeon, Sulfolobus sp. strain 7. J. Bacteriol. 179, 1174-1179. Pubmed

12. Iwasaki, T.,* and Oshima, T. (1997) A stable intermediate product of the archaeal zinc-containing 7Fe ferredoxin from Sulfolobus sp. strain 7 by artificial oxidative conversion. FEBS Lett. 417, 223-226. Pubmed

13. Hori, H, Iwasaki, T., Kurahashi, Y., and Nishino, T.* (1997) Calcium-dependent inactivation of neuronal nitric oxide synthase: evidence for the existence of stabilization/activation factor. Biochem. Biophys. Res. Commun. 234, 476-480. Pubmed

14. Iwasaki, T.,* Suzuki, T., Kon, T., Imai, T., Urushiyama, A., Ohmori, D., and Oshima, T. (1997) Novel zinc-containing ferredoxin family in the thermoacidophilic archaea. J. Biol. Chem. 272, 3453-3458. Pubmed

15. Iwasaki, T.,* Hori, H., Hayashi, Y., and Nishino, T. (1999) Modulation of the remote heme site geometry of recombinant mouse neuronal nitric-oxide synthase by the N-terminal hook region. J. Biol. Chem. 274, 7705-7713. Pubmed

16. Iwasaki, T., Hori, H., Hayashi, Y., Nishino, T.,* Tamura, K., Oue, S., Iizuka, T., Ogura, T., and Esumi, H. (1999) Characterization of mouse nNOS2, a natural variant of neuronal nitric-oxide synthase produced in the central nervous system by selective alternative splicing. J. Biol. Chem. 274, 17559-17566. Pubmed

17. Cosper, N. J., Stålhandske, C. M. V., Iwasaki, H., Oshima, T., Scott, R. A.,* and Iwasaki, T.* (1999) Structural conservation of the isolated zinc site in archaeal zinc-containing ferredoxins as revealed by X-ray absorption spectroscopic analysis and its evolutionary implications. J. Biol. Chem. 274, 23160-23168. Pubmed

18. Hori, H,* Iwasaki, T., Hayashi, Y., Kurahashi, Y., Matsumura, T., and Nishino, T. (1999) Inhibition of neuronal nitric oxide synthase by phosphatidylinositol 4,5-bisphosphate and phosphatidic acid. J. Biochem. 126, 829-837. Pubmed

19. Iwasaki, T.,* Watanabe, E., Ohmori, D., Imai, T., Urushiyama, A., Akiyama, M., Hayashi-Iwasaki, Y., Cosper, N. J., and Scott, R. A. (2000) Spectroscopic investigation of the selective cluster conversion of archaeal zinc-containing ferredoxin from Sulfolobus sp. strain 7. J. Biol. Chem. 275, 25391-25401. Pubmed

20. Iwasaki, T., Okamoto, K., Nishino, T., Mizushima, J., Hori, H., Nishino, T.* (2000) Sequence motif-specific assignment of two [2Fe-2S] clusters in rat xanthine oxidoreductase studied by site-directed mutagenesis. J. Biochem. 127, 771-778. Pubmed

21. Ishii, M.,* Takishita, S., Iwasaki, T., Peerapornpisal, Y., Yoshino, J.-i., Kodama, T., and Igarashi, Y. (2000) Purification and characterization of membrane-bound hydrogenase from Hydrogenobacter thermophilus strain TK-6, an obligately autotrophic, thermophilic, hydrogen-oxidizing bacterium. Biosci. Biotechnol. Biochem. 64, 492-502. J-stage

22. Cosper, N. J., Scott, R. A.,* Hori, H, Nishino, T., and Iwasaki, T.* (2001) X-ray absorption spectroscopic analysis of the high-spin ferriheme site in substrate-bound neuronal nitric-oxide synthase. J. Biochem. 130, 191-198. Pubmed

23. Samoilova, R. I., Kolling, D., Uzawa, T., Iwasaki, T., Crofts, A. R., and Dikanov, S. A.* (2002) The interaction of the Rieske Iron-sulfur protein with occupants of the Qo-site of the bc1 complex, probed by electron spin echo envelope modulation. J. Biol. Chem. 277, 4605-4608. Pubmed

24. Iwasaki, T.,* Kounosu, A., Aoshima, M., Ohmori, D., Imai, T., Urushiyama, A., Cosper, N. J., and Scott, R. A. (2002) Novel [2Fe-2S]-type redox center C in SdhC of archaeal respiratory complex II from Sulfolobus tokodaii strain 7. J. Biol. Chem. 277, 39642-39648. Pubmed

25. Cosper, N. J.,* Eby, D. M., Kounosu, A., Kurosawa, N., Neidle, E. L., Kurtz, D. M. Jr., Iwasaki, T., and Scott, R. A. (2002) Redox-dependent structural changes in archaeal and bacterial Rieske-type [2Fe-2S] clusters. Protein Sci. 11, 2969-2973. Pubmed

26. Suzuki, T., Iwasaki, T., Uzawa, T., Hara, K., Nemoto, N., Kon, T., Ueki, T., Yamagishi, A., and Oshima, T.* (2002) Sulfolobus tokodaii sp. nov. (f. Sulfolobus sp. strain 7), a new member of the genus Sulfolobus isolated from Beppu Hot Springs, Japan. Extremophiles 6, 39-44. Pubmed

27. Yoshimatsu, K., Iwasaki, T., Fujiwara, T.* (2002) Sequence and electron paramegnetic resonance analyses of nitrate reductase NarGH from a denitrifying halophilic euryarchaeote Haloarcula marismortui. FEBS Lett. 516, 145-150. Pubmed

28. Li, Z.,* Shokes, J. E., Kounosu, A., Imai, T., Iwasaki, T., and Scott, R. A. (2003) X-ray absorption spectroscopic analysis of reductive [2Fe-2S] cluster degradation in hyperthermophilic archaeal succinate:caldariellaquinone oxidoreductase subunits. Biochemistry 42, 15003-15008. Pubmed

29. Kounosu, A., Li, Z., Cosper, N. J., Scott, R. A., Imai, T., Urushiyama, A., and Iwasaki, T.* (2004) Engineering a three-cysteine, one-histidine ligand environment into a new hyperthermophilic archaeal Rieske-type [2Fe-2S] ferredoxin from Sulfolobus solfataricus. J. Biol. Chem. 279, 12519-12528. Pubmed

30. Iwasaki, T.,* Kounosu, A., Kolling, D. R. J., Crofts, A. R., Dikanov, S. A., Jin, A., Imai, T., and Urushiyama, A. (2004) Characterization of the pH-dependent resonance Raman transitions of archaeal and bacterial Rieske [2Fe-2S] proteins. J. Am. Chem. Soc. 126, 4788-4789. Pubmed

31. Iwasaki, T.,* Kounosu, A., Uzawa, T., Samoilova, R. I., and Dikanov, S. A.* (2004) Orientation-selected 15N HYSCORE detection of weakly coupled nitrogens around the archaeal Rieske [2Fe-2S] center. J. Am. Chem. Soc. 126, 13902-13903. Pubmed

32. Dikanov, S. A.,* Shubin, A. A., Kounosu, A., Iwasaki, T., and Samoilova, R. I. (2004) A comparative, two-dimensional 14N ESEEM characterization of reduced [2Fe-2S] clusters in hyperthermophilic archaeal high- and low-potential Rieske-type proteins. J. Biol. Inorg. Chem. 9, 753-767. Pubmed

33. Uchiyama, T., Kounosu, A., Sato, T., Tanaka, N., Iwasaki, T.,* and Kumasaka, T.* (2004) Crystallization and preliminary X-ray diffraction studies of the hyperthermophilic archaeal sulredoxin having the unique Rieske [2Fe-2S] cluster environment. Acta Crystallogr. Sect. D 60, 1487-1489. Pubmed

34. Iwasaki, T.,* Kounosu, A., Tao, Y., Li, Z., Shokes, J. E., Cosper, N. J., Imai, T., Urushiyama, A., and Scott, R. A.* (2005) Rational design of a mononuclear metal site into the archaeal Rieske-type protein scaffold. J. Biol. Chem. 280, 9129-9134. Pubmed

35. Iwasaki, T.,* Kounosu, A., Samoilova, R. I., and Dikanov, S. A.* (2006) 15N HYSCORE characterization of the fully deprotonated, reduced form of the archaeal Rieske [2Fe-2S] center. J. Am. Chem. Soc. 128, 2170-2171. Pubmed

36. Iwasaki, T.,* Kounosu, A., Kolling, D. R. J., Lhee, S.-M., Crofts, A. R., Dikanov, S. A., Uchiyama, T., Kumasaka, T., Ishikawa, H., Kono, M., Imai, T., and Urushiyama, A. (2006) Resonance Raman characterization of archaeal and bacterial Rieske protein variants with modified hydrogen bond network around the [2Fe-2S] center. Protein Sci. 15, 2019-2024. Pubmed

37. Iwasaki, T.,* Kounosu, A., Ohmori, D., and Kumasaka, T.* (2006) Crystallization and preliminary X-ray diffraction studies of a hyperthermophilic Rieske protein variant (SDX-triple) with an engineered rubredoxin-like mononuclear iron site. Acta Crystallogr. Sect. F 62, 993-995. Pubmed

38. Iwasaki, T.,* Ohmori, D., Shimizu, N., and Kumasaka, T.* (2007) Crystallization and preliminary X-ray diffraction studies of the ISC-like [2Fe-2S] ferredoxin from Pseudomonas putida JCM 20004. Acta Crystallogr. Sect. F 63, 1014-1016. Pubmed

39. Kounosu, A., Iwasaki, T.,* Baba, S., Hayashi-Iwasaki, Y., Oshima, T., and Kumasaka, T.* (2008) Crystallization and preliminary X-ray diffraction studies of the prototypal homologue of mitoNEET (Tth-NEET0026) from the extreme thermophile Thermus thermophilus HB8. Acta Crystallogr. Sect. F 64, 1146-1148. Pubmed

40. Iwasaki, T.,* Samoilova, R. I., Kounosu, A., and Dikanov, S. A.* (2009) Two-dimensional pulsed electron spin resonance characterization of 15N-labeled archaeal Rieske-type ferredoxin. FEBS Lett. 583, 3467-3472. Pubmed

41. Iwasaki, T.,* Samoilova, R. I., Kounosu, A., Ohmori, D., and Dikanov, S. A.* (2009) Continuous-wave and pulsed EPR characterization of the [2Fe-2S](Cys)3(His)1 cluster in rat mitoNEET. J. Am. Chem. Soc. 131, 13659-13667. Pubmed

42. Kounosu, A., Hasegawa, K., Iwasaki, T.,* and Kumasaka, T.* (2010) Crystallization and preliminary X-ray diffraction studies of hyperthermophilic archaeal Rieske-type ferredoxin (ARF) from Sulfolobus solfataricus P1. Acta Crystallogr. Sect. F 66, 842-845. Pubmed

43. Iwasaki, T.,* Kappl, R.,* Bracic, G., Shimizu, N., Ohmori, D., and Kumasaka, T.* (2011) ISC-like [2Fe-2S] ferredoxin (FdxB) dimer from Pseudomonas putida JCM 20004: structural and electron-nuclear double resonance characterization. J. Biol. Inorg. Chem. 16, 923-935. Pubmed    **PDB code: 3AH7 and interactive 3D complement in Proteopedia

44. Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A.,* Rienstra, C. M.,* and Gennis, R. B.* (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378. Pubmed

45. Iwasaki, T.,* Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A.* (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738. Pubmed

46. Taguchi, A. T., Miyajima-Nakano, Y., Fukazawa, R., Lin, M. T., Baldansuren, A., Gennis, R. B., Hasegawa, K., Kumasaka, T., Dikanov, S. A.,* and Iwasaki, T.* (2018) Unpaired electron spin density distribution across reduced [2Fe-2S] cluster ligands by 13Cβ-cysteine labeling. Inorg. Chem. 57, 741-746. Pubmed

47. Taguchi, A. T.,* Ohmori, D., Dikanov, S. A., and Iwasaki, T.* (2018) g-Tensor directions in the protein structural frame of hyperthermophilic archaeal reduced Rieske-type ferredoxin explored by 13C pulsed electron paramagnetic resonance. Biochemistry 47, 4074-4082. Pubmed

48. Geldenhuys, W. J.,* Long, T. E., Saralkar, P., Iwasaki, T., Nuñez, R. A. A., Nair, R. R., Konkle, M. E., Menze, M. A., Pinti, M. V., Hollander, J. M., Hazlehurst, L. A., and Robart, A. R.* (2019) Crystal structure of the mitochondrial protein mitoNEET bound to a benze-sulfonide ligand. Communications Chemistry 2, Article number: 77. Open access
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Review articles - where possible, include links for you to read them.

49. Nishino, T., Nakanishi, S., Okamoto, K., Mizushima, J., Hori, H., Iwasaki, T., Nishino, T., Ichimori, Y., and Nakagawa, H. (1997) Conversion of xanthine dehydrogenase to oxidase and its role of reperfusion injury. Biochem. Soc. Trans. 25, 783-786. (Review) Pubmed

50. Iwasaki, T.,* and Oshima, T. (2001) Ferredoxin and related enzymes from Sulfolobus. Methods Enzymol. (Hyperthermophilic Enzymes Part C), 334, 3-22. (Review/Methods) Pubmed

51. Iwasaki, T.* (2010) Iron-sulfur world in aerobic and hyperthermoacidophilic archaea Sulfolobus. Archaea Vol. 2010, pii: 842639, 14 pages. doi:10.1155/2010/842639 (Review) Pubmed

52. Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T.,* and Gennis, R. B. (2015) Escherichia coli auxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66. (Review/Methods) Pubmed

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Book section

53. Iwasaki, T., Wakagi, T., Oshima, T., and Matsuura, K. (1994) The absence of protoheme in active respiratory complex II of the thermoacidophilic archaeon, Sulfolobus sp. strain 7. In Flavins and Flavoproteins 1993 (Yagi, K., ed.) pp. 755-758. Walter de Gruyter, Berlin.

54. Nishino, T., Kashima, Y., Okamoto, K., Iwasaki, T., and Nishino, T. (1997) The monomeric form of xanthine dehydrogenase expressed in baculovirus-insect cell system. In Flavins and Flavoproteins 1996 (Stevenson, K. J., Massey, V., and Williams, C. H., Jr., eds.). pp. 843-846. University of Calgury Press, Calgury.

55. Enroth, C., Eger, B., Pai, E. F., Okamoto, K., Iwasaki, T., Nishino, T., Hori, H., and Nishino, T. (1999) Crystal structure and EPR assignment of iron-sulfur centers of xanthine oxidoreductase. In Flavins and Flavoproteins 1999 (Ghisla, S., Kroneck, P., Macheroux, P. and Sund, H., eds.) pp. 783-786, Agency for Scientific Publ., Berlin.

56. Iwasaki, T., Aoshima, M., Kounosu, A., and Oshima, T. (1999) Respiratory complex II from the thermoacidophilic archaeon, Sulfolobus sp. strain 7: genes and protein. In Flavins and Flavoproteins 1999 (Ghisla, S., Kroneck, P., Macheroux, P. and Sund, H., eds.) pp. 779-781, Agency for Scientific Publ., Berlin.

57. Iwasaki, T., Kounosu, A., and Dikanov, S. A. (2002) The [2Fe-2S] cluster in sulredoxin from the thermoacidophilic archaeon Sulfolobus tokodaii strain 7, a novel water-soluble Rieske protein. In EPR in the 21st Century (Kawamori, A., Yamauchi, J., and Ohta, H., eds.), Elsevier Science B.V., Amsterdam, pp. 488-493.
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Review articles in Japanese

58. 堀 弘幸、岩崎 俊雄 「NO合成酵素の構造と機能」 医学のあゆみ、Vol. 188 (No. 13) pp. 1113-1115. 1999. (和文総説)

59. 岩崎 俊雄、鴻巣 麻子 「極限微生物から学ぶ酸素呼吸系獲得への道:エネルギー代謝系機能進化の分子基盤」 極限環境微生物学会誌、No 2 (2). pp. 5-7. 2003. (和文総説)

60. 岩崎 俊雄 「酸素呼吸系の初期機能進化パターン」 極限環境微生物学会誌、No 4 (2). pp. 65-70. 2005.(和文総説)
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Symposium/Workshop - conference abstracts/proceedings 紀要等(査読なし):

1. Iwasaki, T., Wakagi, T., and Oshima, T. (1993) The ferredoxin-dependent red-ox system of the thermoacidophilic archaeon, Sulfolobus sp. strain 7. In International Workshop on Molecular Biology and Biotechnology of Extremophiles and Archaebacteria, pp. 69-70. August 1993, Wako-shi, Saitama, Japan (abstr.).

2. Wakagi, T., Zhang, Q., Iwasaki, T., Fujii, T., Oshima, T., and Matsuzawa, H. (1996) Sulfolobus ferredoxin and related enzymes: structure and function. In Thermophiles '96, pp. 209. August 1996, Georgia, Athens, U.S.A. (abstr.).

3. Iwasaki, T., Okamoto, K., Hori, H., Nishino, T., and Nishino, T. (1997) Xanthine oxidase: EPR assignments of two iron-sulfur clusters. PreICBIC at Okazaki. July 1997, Okazaki, Japan (abstr.).

4. Hori, H, Iwasaki, T., Kurahashi, Y., Hayashi, Y., Umeda, M., and Nishino, T. (1997) Effect of phospholipids on calcium-dependent inactivation of neuronal nitric oxide synthase. In The Biology of Nitric Oxide 6 (Moncada, S., Toda, N., Maeda, H., and Higgs, E. A, eds.). Portland Press, London (abstr.).

5. Iwasaki, T., Hori, H., Hayashi, Y., and Nishino, T. (1997) Heme-binding domain and the threonine mutants of mouse neuronal nitric oxide synthase. J. Inorg. Biochem. 67, 103 (abstr.).

6. Okamoto, K., Iwasaki, T., Nishino, T., Hori, H., Mizushima, J., and Nishino, T. (1997) Properties of xanthine dehydrogenase that lacks iron-sulfur centers. J. Inorg. Biochem. 67, 263 (abstr.).

7. Hori, H, Iwasaki, T., Kurahashi, Y., Hayashi, Y., Umeda, M., and Nishino, T. (1997) Effect of phospholipids on calcium-dependent inactivation of neuronal nitric oxide synthase. Jpn. J. Pharmacol. 75, 26 (abstr.).

8. Iwasaki, T., Imai, T., Urushiyama, A., Ohmori, D., Stålhandske, C., Scott, R. A., and Oshima, T. (1998) "Zinc-containing ferredoxins" from thermoacidophilic archaea: metal centers and evolutionary implication. In The International Conference of Extremophiles 1998, pp. 242-243 (Yokohama, Japan, January 1998) (abstr.).

9. Beniac, D.R., Iwasaki, T., Ottensmeyer, F.P. (1999) In Proc. 26th Ann. Meet. Microsc. Soc. Canada (Corbett, J.M., ed.). Guelph, Ontario, pp. 49-50 (abstr.).

10. Iwasaki, T., and Dikanov, S. A. (1999) Probing the cluster surrounding in sulredoxin from the thermoacidophilic archaeon Sulfolobus sp. strain 7, a novel water-soluble Rieske [2Fe-2S] protein. J. Inorg. Biochem. 74, 113. (abstr.).

11. 岩崎 俊雄 (2001) 「神経型NO合成酵素の構造と機能」. 理研シンポジウム「有機化学反応の新展開(第3回)」講演要旨集、理化学研究所、pp. 13-16 (abstr.).

12. Beniac, D.R., Iwasaki, T., Eger, B., Pai, E., and Ottensmeyer, F.P. (2002) Improvements in single particle cryo-EM: 11Å structure of 290 kDa XDH refined from multiple 3D reconstructions. Microscopy and Microanalysis 8 (Suppl 2), 848-849 (PDF abstract file).

13. Iwasaki, T., Ohmori, D., and Kounosu, A. (2007) Modular structure and assembly of archaeal respiratory complex II. Proceedings of International Symposium on Extremophiles and Their Applications, Vol. 2005, pp. 329 (J-STAGE: http://www.jstage.jst.go.jp/browse/) (abstr.).

14. Iwasaki, T., Kounosu, A., Ohmori, D., Hayashi-Iwasaki, Y., Oshima, T., Dikanov, S. A., Cowart, D. M., Koppolu, U. M. K., Scott, R. A., and Kumasaka, T. (2009) Thermophile "NEET": structure toward functional understanding. (P582) J. Biol. Inorg. Chem. 14 (Suppl 1), S212 (abstr.).

15. Iwasaki, T., Samoilova, R. I., Dikanov, S. A., Miyajima-Nakano, Y., and Kumasaka, T. (2011) Crystal structure and pulsed EPR characterization of hyperthermophilic archaeal Rieske [2Fe-2S] protein (sulredoxin) (#14018). In IXth European Symposium of the Protein Society: Wonders and Disasters of the Protein World, pp. 94. May 2011, Stockholm, Sweden (abstr.).

16. Miyajima-Nakano, Y., Iwasaki, T., Shimizu, N., Kumasaka, T., Kappl, R., Bracic, G., Samoilova, R. I., Baldansuren, A., Lin, M. T., Gennis, R. B., Ohmori, D., and Dikanov, S. A. (2011) Crystal structure and pulsed EPR characterization of the hydrogen bond network around the [2Fe-2S] cluster of Pseudomonas putida ferredoxin (FdxB) involved in the iron-sulfur cluster biosynthesis system (#14023). In IXth European Symposium of the Protein Society: Wonders and Disasters of the Protein World, pp. 95. May 2011, Stockholm, Sweden (abstr.).

17. Iwasaki, T., Miyajima-Nakano, Y., Baldansuren, A., Dikanov, S. A., Hayashi-Iwasaki, Y., Oshima, T., Ohmori, D., and Kumasaka, T. (2011) Structure and metabolome analysis of thermophile "mitoNEET" homolog. (#1107506) 15th International Conference on Bioinorganic Chemistry. August 2011, Vancouver, Canada (abstr.).

18. Hagiuda, E., Miyajima-Nakano, Y., Ohmori, D., Kusano, T., Matsushita, S., Dikanov, S. A., Kumasaka, T., and Iwasaki, T. (2012) Characterization of the extremophile homologs of mitoNEET. (#P117) 9th International Congress on Extremophiles. September 2012, Sevilla, Spain (abstr.).

19. Iwasaki, T., and Kumasaka, T. (2012) Structural basis for metallosulfur recognition of archaeal Rieske protein scaffold. (#P118) 9th International Congress on Extremophiles. September 2012, Sevilla, Spain (abstr.).

20. Fukazawa, R., Lin, M. T., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Choi, S. K., Dikanov, S. A., Gennis, R. B., and Iwasaki, T. (2012) Selective isotope labelling of extremophile metalloproteins. (#P119) 9th International Congress on Extremophiles. September 2012, Sevilla, Spain (abstr.).

21. Hasegawa, K., Matsushita, S., Konosu, A., Baldansuren, A., Fukazawa, R., Dikanov, S. A., Iwasaki, T., and Kumasaka, T. (2012) Active site structure of hyperthermophilic archaeal Rieske-type ferredoxin (ARF). (#P120) 9th International Congress on Extremophiles. September 2012, Sevilla, Spain (abstr.).

22. Matsushita, S., Fukazawa, R., Iwasaki, T., Taguchi, A., Baldansuren, A., and Dikanov, S. A. (2013) 2D pulsed EPR analysis of histidine ligand residue(s) of the thermophile Rieske and mitoNEET type iron-sulfur proteins. (Mz76) 16th International Conference on Bioinorganic Chemistry. July 22-26, 2013, Grenoble, France (abstr.).

23. Iwasaki, T., Miyajima-Nakano, Y., Fukazawa, R., Hagiuda, E., Matsushita, S., Hasegawa, K., Kumasaka, T., Baldansuren, A., and Dikanov, S. A. (2013) Application of 2D pulsed EPR (HYSCORE) in the structural analysis of hydrogen bond network around a biological iron-sulfur cluster. (Mz54) 16th International Conference on Bioinorganic Chemistry. July 22-26, 2013, Grenoble, France (abstr.).

24. Fukazawa, R., Lin, M. T., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Dikanov, S. A., Gennis, R. B., and Iwasaki, T. (2013) Expression host strains for selective isotope labeling of thermophile enzymes. (#AP.P 15) Thermophiles 2013, 12th International Meeting at the University of Regensburg. September 8-13, 2013, Regensburg, Germany (abstr.).

25. Iwasaki, T., Hagiuda, E., Fukazawa, R., Hayashi-Iwasaki, Y., Oshima, T., Dikanov, S. A., Hasegawa, K., and Kumasaka, T. (2013) Thermophile homolog of mitoNEET from Thermus thermophilus HB8. (#FT.O 02) Thermophiles 2013, 12th International Meeting at the University of Regensburg. September 8-13, 2013, Regensburg, Germany (abstr.).

26. Fukazawa, R., Lin, M. T., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Baldansuren, A., Dikanov, S. A., Gennis, R. B., and Iwasaki, T. (2014) A set of Escherichia coli amino acid auxotrophic expression host strains for selectively labelled metalloenzyme research. (P191) J. Biol. Inorg. Chem. 19 (Suppl 2), S845 (abstr.).

27. Iwasaki, T., Hagiuda, E., Fukazawa, R., Hayashi-Iwasaki, Y., Oshima, T., Hasegawa, K., and Kumasaka, T. (2014) Ligand mutagenesis of TthNEET, a thermophile homolog of mitoNEET. (P193) J. Biol. Inorg. Chem. 19 (Suppl 2), S845-846 (abstr.).

28. Taguchi, A. T., Miyajima-Nakano, Y., Fukazawa, R., Baldansuren, A., Samoilova, R. I., Hasegawa, K., Kumasaka, T., Dikanov, S. A., and Iwasaki, T. (2015) Mapping the electron spin distribution in [2Fe-2S] proteins by 13C(beta) cysteine labeling: implications in electron transport pathways. (OL: #A0197) 17th International Conference on Bioinorganic Chemistry. July 20-24, 2015, Beijing, China (abstr.).

29. 岩崎 俊雄 「グルコース感受性に関わる細菌型mitoNEETの生体内レドックス制御システム解析」 長瀬科学技術振興財団 研究報告集、Vol. 26. 2014年度版 pp. 89-97. 2015.(研究報告集・紀要)
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計101件 [国際学会(2+39)件、国内学会(21+39)件] を見る(次のページへ Follow the link)

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社会貢献: 新規アミノ酸要求性大腸菌発現宿主株コレクション
New Escherichia coli amino acid auxotrophic expression strains

EcoliAAmetabmapv5.jpg

See "Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coli auxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66." Pubmed

Amino acid-selective isotope labeling is an extremely powerful method to elucidate specific contributions of particular residues in the reaction mechanisms and/or folding of a target protein by magnetic resonance (e.g., nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR)) and vibrational (e.g., resonance Raman (RR) and Fourier transform infrared (FTIR)) spectroscopies, often aided by the X-ray crystal structure. These techniques can provide detailed information about protein–protein and protein–ligand interactions and dynamics.

One of the most convenient and cost-effective procedures for selective isotope labeling of proteins is to employ amino acid auxotrophic bacteria as the host strains for the overproduction of target proteins. A wild-type Escherichia coli strain has the ability to synthesize all 20 amino acids, whereas an E. coli auxotroph, having an essential gene involved in the biosynthesis of an amino acid disrupted, requires that particular amino acid for growth. However, no suitable auxotrophic strains are commercially available for high-level expression of the foreign genes coding for metalloenzymes from extremophilic archaea and bacteria, because (i) their high-level expression, e.g., in E. coli, often requires extra copies of tRNA genes for the cognate rare codons and (ii) specific growth conditions must be set for effective overproduction of holoproteins in a form suitable for biophysical studies.

To overcome these problems, we have reported the construction of a set of new C43(DE3) and BL21(DE3) auxotrophic expression strains of E. coli designed to facilitate the labeling of either membrane proteins or water-soluble proteins with selected amino acid types enriched with stable isotopes such as 2H, 13C and 15N [Methods Enzymol. 565, 45-66 (2015)]. The use of a suitable auxotrophic expression strain with the corresponding input isotope labeled amino acid(s) in the growth medium ensures high levels of efficiency as well as selectivity in stable isotope labeling, and is expected to solve many selective labeling problems.

*(Almost) All these auxotrophic expression strains of E. coli are made available through Addgene, USA <https://www.addgene.org/Toshio_Iwasaki/>, and LinkIconRIKEN BRC, Japan <https://dnaconda.riken.jp/search/depositor/dep006963.html>.

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