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データを開く
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基本情報
登録情報 | データベース: SASBDB / ID: SASDDT9 |
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![]() | NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) in complex with Ras-related protein Rab 8 (MICAL1-Rab8 complex)
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機能・相同性 | ![]() protein localization to basolateral plasma membrane / establishment of neuroblast polarity / endoplasmic reticulum tubular network organization / establishment of protein localization to endoplasmic reticulum membrane / hippocampal mossy fiber expansion / secretory vesicle / : / F-actin monooxygenase / F-actin monooxygenase activity / NAD(P)H oxidase (H2O2-forming) ...protein localization to basolateral plasma membrane / establishment of neuroblast polarity / endoplasmic reticulum tubular network organization / establishment of protein localization to endoplasmic reticulum membrane / hippocampal mossy fiber expansion / secretory vesicle / : / F-actin monooxygenase / F-actin monooxygenase activity / NAD(P)H oxidase (H2O2-forming) / sulfur oxidation / endoplasmic reticulum tubular network / regulation of regulated secretory pathway / regulated exocytosis / insulin-responsive compartment / NAD(P)H oxidase H2O2-forming activity / polarized epithelial cell differentiation / exocytic vesicle / oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen / cadherin binding involved in cell-cell adhesion / myosin V binding / Golgi to plasma membrane transport / RAB geranylgeranylation / Golgi to plasma membrane protein transport / actin filament depolymerization / RAB GEFs exchange GTP for GDP on RABs / establishment of protein localization to membrane / intermediate filament / endosomal transport / antigen processing and presentation / exocytosis / actin filament bundle assembly / intercellular bridge / vesicle-mediated transport / cytoskeleton organization / FAD binding / cytoplasmic vesicle membrane / axonogenesis / secretory granule membrane / small monomeric GTPase / protein localization to plasma membrane / Translocation of SLC2A4 (GLUT4) to the plasma membrane / adherens junction / actin filament / trans-Golgi network / monooxygenase activity / recycling endosome / small GTPase binding / SH3 domain binding / cellular response to insulin stimulus / recycling endosome membrane / phagocytic vesicle membrane / actin filament binding / GDP binding / actin cytoskeleton / actin binding / G protein activity / Factors involved in megakaryocyte development and platelet production / midbody / cytoskeleton / lysosome / endosome / endosome membrane / ciliary basal body / cilium / Golgi membrane / focal adhesion / Neutrophil degranulation / endoplasmic reticulum membrane / negative regulation of apoptotic process / protein kinase binding / GTP binding / perinuclear region of cytoplasm / Golgi apparatus / signal transduction / extracellular exosome / nucleoplasm / metal ion binding / membrane / plasma membrane / cytosol / cytoplasm 類似検索 - 分子機能 |
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![]() | ![]() タイトル: Human MICAL1: Activation by the small GTPase Rab8 and small-angle X-ray scattering studies on the oligomerization state of MICAL1 and its complex with Rab8. 著者: Alessandro Esposito / Valeria Ventura / Maxim V Petoukhov / Amrita Rai / Dmitri I Svergun / Maria A Vanoni / ![]() ![]() ![]() 要旨: Human MICAL1 is a member of a recently discovered family of multidomain proteins that couple a FAD-containing monooxygenase-like domain to typical protein interaction domains. Growing evidence ...Human MICAL1 is a member of a recently discovered family of multidomain proteins that couple a FAD-containing monooxygenase-like domain to typical protein interaction domains. Growing evidence implicates the NADPH oxidase reaction catalyzed by the flavoprotein domain in generation of hydrogen peroxide as a second messenger in an increasing number of cell types and as a specific modulator of actin filaments stability. Several proteins of the Rab families of small GTPases are emerging as regulators of MICAL activity by binding to its C-terminal helical domain presumably shifting the equilibrium from the free - auto-inhibited - conformation to the active one. We here extend the characterization of the MICAL1-Rab8 interaction and show that indeed Rab8, in the active GTP-bound state, stabilizes the active MICAL1 conformation causing a specific four-fold increase of k of the NADPH oxidase reaction. Kinetic data and small-angle X-ray scattering (SAXS) measurements support the formation of a 1:1 complex between full-length MICAL1 and Rab8 with an apparent dissociation constant of approximately 8 μM. This finding supports the hypothesis that Rab8 is a physiological regulator of MICAL1 activity and shows how the protein region preceding the C-terminal Rab-binding domain may mask one of the Rab-binding sites detected with the isolated C-terminal fragment. SAXS-based modeling allowed us to propose the first model of the free full-length MICAL1, which is consistent with an auto-inhibited conformation in which the C-terminal region prevents catalysis by interfering with the conformational changes that are predicted to occur during the catalytic cycle. |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
-モデル
モデル #2179 | ![]() タイプ: mix / ダミー原子の半径: 1.90 A / カイ2乗値: 1.28 / P-value: 0.006548 ![]() |
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モデル #2180 | ![]() タイプ: mix / ダミー原子の半径: 1.90 A / カイ2乗値: 1.39 / P-value: 0.000409 ![]() |
モデル #2181 | ![]() タイプ: mix / ダミー原子の半径: 1.90 A / カイ2乗値: 1.50 ![]() |
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試料
![]() | 名称: NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) in complex with Ras-related protein Rab 8 (MICAL1-Rab8 complex) 試料濃度: 10 mg/ml / Entity id: 1192 / 1194 |
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バッファ | 名称: 20 mM Hepes/NaOH, pH 7.5, 50 mM NaCl, 2 mM MgCl2, 2 mM DTT pH: 7.5 / コメント: Mical1:Rab8 |
要素 #1192 | 名称: Mical1 / タイプ: protein / 記述: [F-actin]-monooxygenase MICAL1 (monomer) / 分子量: 117.93 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: Q8TDZ2 配列: MASPTSTNPA HAHFESFLQA QLCQDVLSSF QELCGALGLE PGGGLPQYHK IKDQLNYWSA KSLWTKLDKR AGQPVYQQGR ACTSTKCLVV GAGPCGLRVA VELALLGARV VLVEKRTKFS RHNVLHLWPF TIHDLRALGA KKFYGRFCTG TLDHISIRQL QLLLLKVALL ...配列: MASPTSTNPA HAHFESFLQA QLCQDVLSSF QELCGALGLE PGGGLPQYHK IKDQLNYWSA KSLWTKLDKR AGQPVYQQGR ACTSTKCLVV GAGPCGLRVA VELALLGARV VLVEKRTKFS RHNVLHLWPF TIHDLRALGA KKFYGRFCTG TLDHISIRQL QLLLLKVALL LGVEIHWGVT FTGLQPPPRK GSGWRAQLQP NPPAQLANYE FDVLISAAGG KFVPEGFKVR EMRGKLAIGI TANFVNGRTV EETQVPEISG VARIYNQSFF QSLLKATGID LENIVYYKDD THYFVMTAKK QCLLRLGVLR QDWPDTNRLL GSANVVPEAL QRFTRAAADF ATHGKLGKLE FAQDAHGQPD VSAFDFTSMM RAESSARVQE KHGARLLLGL VGDCLVEPFW PLGTGVARGF LAAFDAAWMV KRWAEGAESL EVLAERESLY QLLSQTSPEN MHRNVAQYGL DPATRYPNLN LRAVTPNQVR DLYDVLAKEP VQRNNDKTDT GMPATGSAGT QEELLRWCQE QTAGYPGVHV SDLSSSWADG LALCALVYRL QPGLLEPSEL QGLGALEATA WALKVAENEL GITPVVSAQA VVAGSDPLGL IAYLSHFHSA FKSMAHSPGP VSQASPGTSS AVLFLSKLQR TLQRSRAKEN AEDAGGKKLR LEMEAETPST EVPPDPEPGV PLTPPSQHQE AGAGDLCALC GEHLYVLERL CVNGHFFHRS CFRCHTCEAT LWPGGYEQHP GDGHFYCLQH LPQTDHKKEG SDRGPESPEL PTPSENSMPP GLSTPTASQE GAGPVPDPSQ PTRRQIRLSS PERQRLSSLN LTPDPEMEPP PKPPRSCSAL ARHALESSFV GWGLPVQSPQ ALVAMEKEEK ESPFSSEEEE EDVPLDSDVE QALQTFAKTS GTMNNYPTWR RTLLRRAKEE EMKRFCKAQT IQRRLNEIEA ALRELEAEGV KLELALRRQS SSPEQQKKLW VGQLLQLVDK KNSLVAEEAE LMITVQELNL EEKQWQLDQE LRGYMNREEN LKTAADRQAE DQVLRKLVDL VNQRDALIRF QEERRLSELA LGTGAQG |
要素 #1194 | 名称: Rab8 / タイプ: protein / 記述: Ras-related protein 8 / 分子量: 20.171 / 分子数: 1 / 参照: UniProt: P61026 配列: GHMAKKTYDL LFKLLLIGDS GVGKTCVLFR FSDDAFNTTF ISTIGIDFKI KTVELQGKKI KLQIWDTAGQ ERFHTITTSY YRGAMGIMLV YDITNGKSFE NISKWLRNID EHANEDVERM LLGNKCDMDD KRVVPKGKGE QIAREHGIRF FETSAKANIN IEKAFLTLAE DILRKTP |
-実験情報
ビーム | 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany ![]() | |||||||||||||||||||||
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検出器 | 名称: Pilatus 2M | |||||||||||||||||||||
スキャン | 測定日: 2017年6月20日 / 保管温度: 23 °C / セル温度: 10 °C / 照射時間: 1 sec. / フレーム数: 3600 / 単位: 1/nm /
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距離分布関数 P(R) |
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結果 | コメント: 1:1 Complex between Mical1 and Rab8. Note: The protein in the SAXS experiments is Rab8, however the protein used for modelling is Rab10.
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