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基本情報
登録情報 | データベース: SASBDB / ID: SASDDP3 |
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![]() | N-propargyl glycine-Inactivated Proline utilization A from Bradyrhizobium diazoefficiens (formerly Bradyrhizobium japonicum) collected by SEC-SAXS
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機能・相同性 | ![]() proline dehydrogenase / proline dehydrogenase activity / L-glutamate gamma-semialdehyde dehydrogenase / 1-pyrroline-5-carboxylate dehydrogenase activity / proline catabolic process to glutamate / proline biosynthetic process / cytoplasmic side of plasma membrane / DNA-binding transcription factor activity / nucleotide binding / DNA binding / identical protein binding 類似検索 - 分子機能 |
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![]() | ![]() タイトル: Redox Modulation of Oligomeric State in Proline Utilization A. 著者: David A Korasick / Ashley C Campbell / Shelbi L Christgen / Srinivas Chakravarthy / Tommi A White / Donald F Becker / John J Tanner / ![]() 要旨: Homooligomerization of proline utilization A (PutA) bifunctional flavoenzymes is intimately tied to catalytic function and substrate channeling. PutA from Bradyrhizobium japonicum (BjPutA) is unique ...Homooligomerization of proline utilization A (PutA) bifunctional flavoenzymes is intimately tied to catalytic function and substrate channeling. PutA from Bradyrhizobium japonicum (BjPutA) is unique among PutAs in that it forms a tetramer in solution. Curiously, a dimeric BjPutA hot spot mutant was previously shown to display wild-type catalytic activity despite lacking the tetrameric structure. These observations raised the question of what is the active oligomeric state of BjPutA. Herein, we investigate the factors that contribute to tetramerization of BjPutA in vitro. Negative-stain electron microscopy indicates that BjPutA is primarily dimeric at nanomolar concentrations, suggesting concentration-dependent tetramerization. Further, sedimentation-velocity analysis of BjPutA at high (micromolar) concentration reveals that although the binding of active-site ligands does not alter oligomeric state, reduction of the flavin adenine dinucleotide cofactor results in dimeric protein. Size-exclusion chromatography coupled with multiangle light scattering and small-angle x-ray scattering analysis also reveals that reduced BjPutA is dimeric. Taken together, these results suggest that the BjPutA oligomeric state is dependent upon both enzyme concentration and the redox state of the flavin cofactor. This is the first report, to our knowledge, of redox-linked oligomerization in the PutA family. |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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-モデル
モデル #1805 | ![]() タイプ: atomic / カイ2乗値: 2.9816132533 ![]() |
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モデル #1795 | ![]() タイプ: atomic / カイ2乗値: 1.64908802698 ![]() |
モデル #1796 | ![]() タイプ: atomic / カイ2乗値: 1.64908802698 ![]() |
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試料
![]() | 名称: N-propargyl glycine-Inactivated Proline utilization A from Bradyrhizobium diazoefficiens (formerly Bradyrhizobium japonicum) collected by SEC-SAXS |
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バッファ | 名称: 50 mM Tris, 50 mM NaCl, 0.5 mM TCEP, 5% (v/v) glycerol pH: 7.8 |
要素 #976 | 名称: BjPutA / タイプ: protein / 記述: Bifunctional protein PutA / 分子量: 107.558 / 分子数: 2 由来: Bradyrhizobium diazoefficiens (strain JCM 10833 / IAM 13628 / NBRC 14792 / USDA 110) 参照: UniProt: Q89E26 配列: GHMPNIPPPF TAPYAPDDAE IAARLLPASH LSPPQEARIH RTATRLIEAI RKRDDRLGGV EDMLREFALS TKEGLALMVL AEALLRVPDA RTADQFIEDK LGEGDFIHHE TKSTAFLVNA SAWALGLSAR VIQPGETPDG TIGRLVKRLG APAVRTATRQ AMRLMGNHFV ...配列: GHMPNIPPPF TAPYAPDDAE IAARLLPASH LSPPQEARIH RTATRLIEAI RKRDDRLGGV EDMLREFALS TKEGLALMVL AEALLRVPDA RTADQFIEDK LGEGDFIHHE TKSTAFLVNA SAWALGLSAR VIQPGETPDG TIGRLVKRLG APAVRTATRQ AMRLMGNHFV LGETIEQALE RGKPRSGQKT RYSFDMLGEG ARTAADARRY FDAYASAIET IGKAAGNHAL PDRPGISVKL SALHPRFEAI SRARVMVELV PQLLDLAQRA KAHDLNFTVD AEEADRLELS LDVIAATLAD PSLKGWDGFG LAIQAYQKRA SAVIDYVDAL ARAHDRKLMV RLVKGAYWDT EIKRAQERGL DGYPVFTRKA MTDLNYVACA SKLLALRPRI FPQFATHNAL TVATVLEMAE GSSGFEFQRL HGMGEALYEQ LAKDHADIAY RTYAPVGSHR DLLAYLVRRL LENGANSSFV AQAADYRVPV PALLQRPADA IVRPQAAAHP RIPLPCDLFA PERRNSRGVE FGARTALDQL LTDVKAETGD LKPIADATPD QAHAAVAAAR AGFAGWSRTP AGIRAAALEQ AAHLLESRSA HFIALLQREG GKTLDDALSE LREAADFCRY YAAQGRKLFG SETAMPGPTG ESNALTMRGR GVFVAISPWN FPLAIFLGQV TAALMAGNSV VAKPAEQTPR IAREAVALLH EAGIPKSALY LVTGDGRIGA ALTAHPDIAG VVFTGSTEVA RSINRALAAK DGPIVPLIAE TGGINAMIAD ATALPEQVAD DVVTSAFRSA GQRCSALRLL FVQEDVADRM IEMVAGAARE LKIGDPSDVA THVGPVIDVE AKQRLDAHIA RMKTEARLHF AGPAPEGCFV APHIFELTEA GQLTEEVFGP ILHVVRYRPE NLERVLRAIE RTGYGLTLGV HSRIDDSIEA IIDRVQVGNI YVNRNMIGAV VGVQPFGGNG LSGTGPKAGG PHYLARFATE QTVTINTAAA GGNAALLAGE E |
-実験情報
ビーム | 設備名称: Advanced Photon Source (APS) BioCAT 18ID / 地域: Argonne, IL / 国: USA ![]() | ||||||||||||||||||||||||||||||
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検出器 | 名称: Pilatus 100K / Pixsize x: 172 mm | ||||||||||||||||||||||||||||||
スキャン | 測定日: 2017年7月16日 / セル温度: 22 °C / 単位: 1/A /
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距離分布関数 P(R) |
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結果 |
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