䠇䌝䠋㻃䝃䝷䝕䜳䠅䠂䠂䠂䝛䝱䜼䜫䜳䝌㻃 ୯ ᶭ㛭ᠺᯕ ሒ 㻃 ࢡ よᯊ ࢡ 㸝 ㅨ 㸞 ୯ ᶭ㛭 ኬ㜨ኬᏕኬᏕ㝌 Ꮥ ಲ ක ᠺ 㸦㸣ᖲᠺ 1㸬ᖳ 10 ᮆ ࡄ ࢡ ධమ ᴏゝ ᑊࡌ ᴏ 㐅ᤎ Ἓ ᴣさ ࡗ Ṟ ሾ ᮮ ㅨ ࢰ ࢠ ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ ይ Ⳟ ࢰ ࢠ ࡗ ኬ㔖 ᬏよ

Size: px

Start display at page:

Download "䠇䌝䠋㻃䝃䝷䝕䜳䠅䠂䠂䠂䝛䝱䜼䜫䜳䝌㻃 ୯ ᶭ㛭ᠺᯕ ሒ 㻃 ࢡ よᯊ ࢡ 㸝 ㅨ 㸞 ୯ ᶭ㛭 ኬ㜨ኬᏕኬᏕ㝌 Ꮥ ಲ ක ᠺ 㸦㸣ᖲᠺ 1㸬ᖳ 10 ᮆ ࡄ ࢡ ධమ ᴏゝ ᑊࡌ ᴏ 㐅ᤎ Ἓ ᴣさ ࡗ Ṟ ሾ ᮮ ㅨ ࢰ ࢠ ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ ይ Ⳟ ࢰ ࢠ ࡗ ኬ㔖 ᬏよ"

蚁稚坦蒙
5 years ago
Views:

䠇䌝䠋㻃䝃䝷䝕䜳䠅䠂䠂䠂䝛䝱䜼䜫䜳䝌㻃 ୯ ᶭ㛭ᠺᯕ ሒ 㻃 ࢡ よᯊ ࢡ 㸝 ㅨ 㸞 ୯ ᶭ㛭 ኬ㜨ኬᏕኬᏕ㝌 Ꮥ ಲ ක ᠺ 㸦㸣ᖲᠺ 1㸬ᖳ 10 ᮆ ࡄ ࢡ ධమ ᴏゝ ᑊࡌ ᴏ 㐅ᤎ Ἓ ᴣさ ࡗ Ṟ ሾ ᮮ ㅨ ࢰ ࢠ ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ ይ Ⳟ ࢰ ࢠ ࡗ ኬ㔖 ᬏよᯊ ຝ ࡂ ࡒ 㸡୯ ᶭ㛭

Ț ࢡ ࢰ ࢨࢪ ᠺ㓕 ᅒ A-1 㸯 Ț-ࢡ ࢰ ࢨࢪ ᠺ㓕 (GCS) 㸡ࢡ ࢰࢲ ᠺࡌ 㝷 ᚂ ṹ㝭 ཬᚺ㸝ࢡ ࢰ 㓗㸠ࢨࢪ 㸞り፳ࡊ㸡మහ ࢡ ࢰࢲ ಕࡖ ࠊࡊ ࡊ㸡ᐞ ཋ 㸡ࢡ ࢰࢲ 㓗 ࢪ ࢪ㜭ᚒࢨࢪ ᝇ ࡌ ࡆ ࡖ ኣ ᛮ ᚋࡊ ࠊኬ Ⳟ ᮮ GCS ᵋ 㐼 ࢡ 々 మ ᬏᵋ Ử ࡊࡒ ᯕ㸡ᚉᮮ 㒼า ῼ

3- ࢨ㓏㓗 ࢣ ࢭ㸯 ᑺ 㸡㓗 ㅨ ஸ㐅 ࡋ ࢢ మ ࢨ ࢨࢪ ࡊ ណㆉ㝸ᐐ ᣅࡂࡒ 㸡 ୯ࢢ మ ᗐ ࢰ ࡌ ᚪさ ࠊ3- ࢨ㓏㓗 ࢣ ࢭ 㸡 3- ࢨ㓏㓗 ࢬ 㓉㓗㸝 ࡍ ࢢ మ㸞 Ề ཬᚺ り፳ࡌ 㓕㸡 ᑺ チ ࡊ ࡈ ࠊᮇ㓕 ᬏᵋ 㓕 NADH ࡊࡒ 㓕 ࡗ Ử ࡊࡒࠊ A-3.

ን 㐅ࡌ ࡆ SerR Ὡ ᛮ ࡌ ࡆ ࡖࡒࠊࡆ ሒ 㸡SerR 㜴ᐐ ୯ ንᛮ ᝀ ᑊࡌ ࡊ 㛜 ࡌ 㐠㛜ࡄࡒࠊ A-4.

1 䠇䌝䠋㻃䝃䝷䝕䜳䠅䠂䠂䠂䝛䝱䜼䜫䜳䝌㻃 ୯ ᶭ㛭ᠺᯕ ሒ 㻃 ࢡ よᯊ ࢡ 㸝 ㅨ 㸞 ୯ ᶭ㛭 ኬ㜨ኬᏕኬᏕ㝌 Ꮥ ಲ ක ᠺ 㸦㸣ᖲᠺ 1㸬ᖳ 10 ᮆ ࡄ ࢡ ධమ ᴏゝ ᑊࡌ ᴏ 㐅ᤎ Ἓ ᴣさ ࡗ Ṟ ሾ ᮮ ㅨ ࢰ ࢠ ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ ይ Ⳟ ࢰ ࢠ ࡗ ኬ㔖 ᬏよᯊ ຝ ࡂ ࡒ 㸡୯ ᶭ㛭 ᪃ス ࢡ ධమ Ὡ ࡊࡗࡗ㸡 ᣈⅤ ᖲ ࡊ ᵋ ᶭ よᯊ 㐅 ࠊ ᵾ よᯊᩐ 㐡ᠺࡊ ࡀ 㸡ࡈ ᚋ ࡒᠺᯕ セ ࡒ ᴏ ࡗࡄ ງ ງ ࡖ ࠊᵋ ᶭ よᯊ ᠺᯕ ࡊ 㸡ḗ ᣪࡅ ࡆ ࡀ ࠊ A. Ꮥ㛭 ࢰ ࢠ A-1. Ț ࢡ ࢰ ࢨࢪ ᠺ㓕 ᅒ A-1 㸯 Ț-ࢡ ࢰ ࢨࢪ ᠺ㓕 (GCS) 㸡ࢡ ࢰࢲ ᠺࡌ 㝷 ᚂ ṹ㝭 ཬᚺ㸝ࢡ ࢰ 㓗㸠ࢨࢪ 㸞り፳ࡊ㸡మහ ࢡ ࢰࢲ ಕࡖ ࠊࡊ ࡊ㸡ᐞ ཋ 㸡ࢡ ࢰࢲ 㓗 ࢪ ࢪ㜭ᚒࢨࢪ ᝇ ࡌ ࡆ ࡖ ኣ ᛮ ᚋࡊ ࠊኬ Ⳟ ᮮ GCS ᵋ 㐼 ࢡ 々 మ ᬏᵋ Ử ࡊࡒ ᯕ㸡ᚉᮮ 㒼า ῼ ࡖࡒࢨࢪ ᵕᘟ ཬᚺᶭᵋ ᥆ ྊ ࡖࡒࠊGCS Ὡᛮ ຝᯕ ᢒโࡌ スゝ ࡀ ࠉ ຝ ᢘᐞ ཋ ᢘ 㛜 ࡗ ᙽࡈ 㸡ぞ㜴ᐐ モ㥺㛜ጙࡊ ࠊ A ࢨ㓏㓗 ࢣ ࢭ㸯 ᑺ 㸡㓗 ㅨ ஸ㐅 ࡋ ࢢ మ ࢨ ࢨࢪ ࡊ ណㆉ㝸ᐐ ᣅࡂࡒ 㸡 ୯ࢢ మ ᗐ ࢰ ࡌ ᚪさ ࠊ3- ࢨ㓏㓗 ࢣ ࢭ 㸡 3- ࢨ㓏㓗 ࢬ 㓉㓗㸝 ࡍ ࢢ మ㸞 Ề ཬᚺ り፳ࡌ 㓕㸡 ᑺ チ ࡊ ࡈ ࠊᮇ㓕 ᬏᵋ 㓕 NADH ࡊࡒ 㓕 ࡗ Ử ࡊࡒࠊ A-3. ᆵࢬ ࢬ ࢭ (ᅒ A-3)㸯 D-ࢬ 㸡୯ᯙ NMDA ࢬ ࢰ ࡌ ࢥ ࢪ 㸡㧏ḗᶭ ࡖ 㔔さ ᙲ ᯕࡒࡊ ࠊࢬ ࢬ ࢭ (SerR) L-ࢬ D-ࢬ ᠺࡌ 㓕㸡 ᅂ㸡ฦ 㓕ẍ SerR ᬏᵋ Ử ࡊࡒࠊࡐ Ὡᛮ㒂న 㸡り፳ᇱ ࢨ ࢰ B6 㛣 D- ᯣᶣࡊࡒಞ㣥ᵋ ᣚࡖ ࡒࠊ ࡒ㸡ATP open-closed ᵋ ን 㐅ࡌ ࡆ SerR Ὡ ᛮ ࡌ ࡆ ࡖࡒࠊࡆ ሒ 㸡SerR 㜴ᐐ ୯ ንᛮ ᝀ ᑊࡌ ࡊ 㛜 ࡌ 㐠㛜ࡄࡒࠊ A-4. Ⳟ ᢘ ฝ ࢪ ࢰ (ᅒ A-4)㸯㝌හវ ཋᅄⳞ ᢘ ᅏ㞬 Ⳟ 㸡 ᢘ ฝ ࢪ ࢰ ࡌ ࡆ ࡖ ኣ ᛮ ࡌࠊࡆ ฝ 㸡 አ ᆵ OprM㸡 አ හ 㛣 న ࡌ MexA㸡 හ ᆵ మ MexB 3 ࡗ ࡖ ᵋᠺࡈ ࠊ ᅂ㸡OprM MexA ᬏᵋ ࡊࡒࠊOprM 㔖మ አ ࡌ ș 㒂ฦ 㛣㝵 ฝࡌ Ș 㒂ฦ 㸡 ὕ ᙟᠺࡊ ࡒࠊMexA 3 ࡗ 㸡OprM MexB ࡌ ฦᏄࢠ ᵋ ࡖࡒࠊࡆ ᵋ ฝ 㜴ᐐ 㛜 ࡀ 㸡ᑛ㔖 ష ᑛ ຝᯕ ᙽ ࡀ ࡓࡄ ࡂ㸡ᚉᮮ ᢘ ྊ 㸡ኣࡂ Ⅴ ᙽࡈ ࠊ Ț ࢡ ࢰ ࢨࢪ ᠺ㓕 ᆵࢬ ࢬ ࢭ OprM + MexA 々 మ (A-4) (A-1) (A-3) (MexB ᵋ ࡎࡒ) A-5. ࡐ 㸯 ࢦ ࢯ ᆵ ฦᒪ㙈㓗 ᇱ 㓕 ࢲ 㸝㸞々 మ㸡 ࢨ Ⳟ ࢰẐ Ia ᠺฦ㸡ࢬ ࢪⳞࢪ ࢥ ࢭ㸝ȘẐ 㸞㸡ᑺ㓗㓗㓕 ࢭ㸝チ㸞㸡 ࢪ ࢯ ࢰ ࢠ Dsg1 ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ

1 O- 85-100 O- B-4. 2- -5- -trna L- (B-1) NAD (B-2) C. C-1.

2 B. B-1. L- ( B-1) L- B-2. NAD ( B-2) (GluDH) GluDH GluDH GluDH NAD NADP GluDH B-3. 1 O O- B trna L- (B-1) NAD (B-2) C. C-1. N- -L- ( C-1) N- - NADPH N- -L- NADPH NADPH N- -L- L- C ( C-2) (S)-2- (S2-CPA) 2- (2-CAA) NADPH 2-CAA S2-CPA

L- 17 4 17 10 14 4 17 3 (1) PDB 1 45 118 (2) PDB 1

3 C-3. (R-CN) (R-CHN-OH) C-4. N- N- -L- (C-1) 2- (C-2) D. D-1. D-2. L- L- 10% D-3. L (1) PDB (2) PDB 1 (3) PDB (1)

4 (2) (3) (4) DNA 1-B-1 DERA N- -L- 1-C-1 heterofructosaccharide 1-B-2 DERA 1-B-4 L- 1-B-1 N- N- -L- N- -L- 1-C-1 AIST 1-B-3 4-(2)-(3) 1) DNA 6-N ) , GDP- D- GMP 17 3) N- -, D-

5 1) N N N N 2) Streptomyces 3) 4) (1) (2) (3) (3) 5) ab initio ACORN, SHARP, warp 1.9 Å ab initio 1.6 Å directed evolution X 16 1) 2) 3) 4) ) 14, 15, 16 17

6 (17 ) 1) 2) 3) SPring-8 PF X SPring-8 4) 5) (

7 Ikeda, H., Ishikawa, J., Hanamoto, A., Shinose, M., Kikuchi, H., Shiba, T., Sakaki, Y., Hattori, M., and Omura, S., Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis, Nature Biotechnol. 21, (2003). Herai, S., Hashimoto, Y., Higashibata, H., Maseda, H., Ikeda, H., Omura, S., and Kobayashi, M., Hyper-inducible expression system for streptomycetes, Proc. Natl. Acad. Sci. USA., 102, (2004). Kuwata, K., Matumoto, T., Cheng, H., Nagayama, K., James, TL., and Roder, H., NMR-detected hydrogen exchange and molecular dynamics simulations provide structural insight into fibril formation of prion protein fragment , Proc. Natl. Acad. Sci. U.S.A., 100 (25), (2003). Tamura, T., Yamamoto, S., Takahata, M., Sakaguchi, H., Tanaka, H., Stadtman, TC., and Inagaki, K., Selenophosphate synthetase genes Sps1 and Sps2 from lung adenocarcinoma cells; Sps1 for recycling L-selenocysteine and Sps2 for selenite assimilation., Proc. Natl. Acad. Sci. USA, 101, (2004). Mitsui, K., Ochi, F., Nakamura, N., Doi, Y., Inoue, H., and Kanazawa, H., A novel membrane protein capable of binding the Na+/H+ antiporter (Nha1p) enhances the salinity-resistant cell growth of Saccharomyces cerevisiae, J. Biol. Chem., 279, (2004). Kuwabara, N., Inoue, H., Tsuboi, Y., Nakamura, N., and Kanazawa, H., The Fourth Transmembrane Segment of Helicobacter pylori Na+/H+ Antiporter NhaA Faces a Water-filled Channel Required for Ion Transport, J. Biol. Chem. 279, (2004). Nakamura, N., Tanaka, S., Teko, Y., and Kanazawa H., Four Na+/H+ exchanger isoforms are distributed to Golgi and post-golgi compartments and are involved in organelle ph regulation. J. Biol. Chem. 280, (2005). Naoe, Y., Arita,K., Hashimoto, H., Kanazawa, H., Sato, M., and Shimizu, T., Structural Characterization of Calcineurin B Homologous Protein 1, J. Biol. Chem., 280, (2005).

8 Konishi, K., Ishida, K., Oinuma, K-I., Ohta, T., Hashimoto, Y., Higashibata, H., Kitagawa, T. & Kobayashi, M. "Identification of crucial histidines involved in carbon-nitrogen triple bond synthesis by aldoxime dehydratase" J. Biol. Chem., 279, (2004). Herai, S., Hashimoto, Y., Higashibata, H., Maseda, H., Ikeda, H., Omura, S. & Kobayashi, M. "Hyper-inducible expression system for streptomycetes" Proc. Natl. Acad. Sci. USA, 101, (2004). Fukatsu, H., Hashimoto, Y., Goda, M., Higashibata, H. & Kobayashi, M. "Amine-synthesizing enzyme, N-substituted formamide deformylase: Screening, purification, characterization and gene cloning" Proc. Natl. Acad. Sci. USA, 101, (2004). Oinuma, K-I., Kumita, H., Ohta, T., Konishi, K., Hashimoto, Y., Higashibata, H., Kitagawa, T., Shiro, Y. & Kobayashi, M. Stopped-flow spectrophotometric and resonance Raman analyses of aldoxime dehydratase involved in carbon-nitrogen triple bond synthesis FEBS Lett., 579, (2005). Nishimasu, H., Fushinobu, S., Shoun, H., and Wakagi, T., Structure, 12 (6) (2004), PDB ID: 1UMG Mizuno, M., Tonozuka, T., Suzuki, S., Uotsu-Tomita, R., Kamitori, S., Nishikawa, A. & Sakano, Y., Structural insights into substrate specificity and function of glucodextranase. J. Biol. Chem., 279, (2004). PBID:1UG9,1ULV. Hayashi, F., Itoh N., Uzumaki, T., Iwase,R., Tsuchiya, Y., Yamakawa, H., Morishita M., Onai K., Itoh, S., and Ishiura, M., Roles of two ATPase-motif-containing domains in cyanobacerial circadian clock protein KaiC., J. Biol. Chem., 279(50), (2004). Tsukatani, R., Miyamoto, Itoh, S., and Oh-oka, H., Function of a PscD subunit in a homodimeric reaction center complex of the photosynthetic green sulfur bacterium Chlorobium tepidum studied by insertional gene inactivation: Regulation of energy transfer and ferredoxin-mediated NADP+ reduction on the cytoplasmic side, J. Biol. Chem., 279(49), (2004). Muramatsu H, Mihara H, Kakutani R, Yasuda M, Ueda M, Kurihara T, Esaki N, The putative malate/lactate dehydrogenase from Pseudomonas putida is an NADPH-dependent Delta(1)-piperideine-2-carboxylate/Delta(1)-pyrroline-2-carboxylate reductase involved in the catabolism of D-lysine and D-proline, J. Biol.

9 Chem. 280, (2005). Mihara H, Kato S-i, Lacourciere GM, Standman TC, Kennedy RAJD, Kurihara T, Tokumoto U, Takahashi Y, Esaki N, The iscs gene is essential for the biosynthesis of 2-selenouridine in trna and the selenocysteine containing formate dehydrogenase H, Proc. Natl. Acad. Sci. USA 99, (2002). Hayashi H, Mizuguchi H, Miyahara I, Nakajima Y, Hirotsu K, Kagamiyama H, Conformational change in aspartate aminotransferase on substrate binding induces strain in the catalytic group and enhances catalysis, J. Biol. Chem. 278(11), (2003). PDB ID: 1IX6, 1IX7, 1IX8. Tsuge H., Kawakami R., Sakuraba H., Ago H., Miyano M., Aki K., Katunuma N., Ohshima T. L-proline dehydrogenase complex from Pyrococcus horikoshii, J. Biol. Chem. 280, ( 2005). PDB ID 1Y56. Crystal structure of a novel FAD-, FMN-, and ATP-containing Sakuraba H., Tsuge H., Yoneda K., Katunuma N., Ohshima T. J. Biol. Chem. 280, ( 2005 ). PDB ID 1WZU. Crystal structure of the NAD biosynthetic enzyme quinolinate synthase Hibi T., Nii H., Nakatsu T., Kimura A., Kato H., Hiratake J., and Oda J., Crystal structure of -gltutamyl-cysteine synthetase: Insights into the mechanism of catalysis by a key enzyme for glutathione homeostasis, Proc. Natl. Acad. Sci. USA, 101, (2004), PDB ID: 1V4G, 1VA6. Akama, H., Kanemaki, M., Tsukihara, T., Nakagawa, A., and Nakae,T., Preliminary crystallographic analysis of the antibiotic-discharge outer membrane lipoprotein, OprM, of Pseudomonas aeruginosa with an exceptionally long unit cell and complex lattice structure, Acta Cryst., F61, (2005). PDB ID: 1WP1. Shibata N, Mori K, Hieda N, Higuchi Y, Yamanishi M, Toraya T, Release of a damaged cofactor from a coenzyme B12-dependent enzyme: X-ray structures of diol dehydratase-reactivating factor, Structure, in press (2005). PDB ID: 2D0O, 2D0P. Kohashi, PY., Kumagai, T., Matoba, Y., Yamamoto, A., Maruyama, M.and Sugiyama, M. An efficient method for the overexpression and purification of the active tyrosinase from Streptomyces castaneoglobisporus. Protein Expr. Purif (2004).

10 Noda, M., Kawahara, Y., Ichikawa, A., Matoba, Y., Matsuo, H., Lee, DG., Kumagai, T. and Sugiyama, M. Self-protection mechanism in D-cycloserine-producing Streptomyces lavendulae. Gene cloning, characterization, and kinetics of its alanine racemase and D-alanyl-D-alanine ligase, which are target enzymes of D-cycloserine. J. Biol. Chem (2004).

untitled

untitled 1 461 Ṟ ሾ ᮮ ㅨ ࢰ ࢠ ࡗ 㸡ᵋ ᶭ よᯊ ࡖࡒࠊ ይ Ⳟ ࢰ ࢠ ࡗ ୯ ᶭ㛭 1000 㢦 w ࢰ ࢠ ࢪ ᵋ ኬ㔖㞗୯ 㸡 ࢪ 300 㢦௧ ࢰ ࢠ ࢦ ᶭ㛭 ࡊ よᯊ ౩ࡌ ࡆ 㸡ࢡ ධ మ ࡊ ຝ ࡂスങ ㈠ᮞ Ὡ ࡊࡒࠊ ࡒ ᣈⅤ ᖲ ࡊ ㅨ ࢰ ࢠ ࡗ ᵋ ᶭ よᯊ 㐅 ࡒࠊ ᵋ ᶭ よᯊ ᠺᯕ ࡊ 㸡ḗ ᣪࡅ ࡆ ࡀ ࠊ & Ꮥ㛭 ฦ㔕 ࢰ ࢠ & Ț ࢡ ࢰ ࢨࢪ ᠺ㓕