SASDDS9: NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (mo...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDDS9

試料

NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) (Truncated MOCHLIM construct)

• [F-actin]-monooxygenase MICAL1 (MoChLim) (protein), MoChLim, Homo sapiens

機能・相同性

分子機能:

hippocampal mossy fiber expansion / NADPH oxidase H202-forming activity / F-actin monooxygenase / F-actin monooxygenase activity / NAD(P)H oxidase (H2O2-forming) / sulfur oxidation / regulation of regulated secretory pathway / NAD(P)H oxidase H2O2-forming activity / 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 / actin filament depolymerization / intermediate filament / intercellular bridge / actin filament bundle assembly / cytoskeleton organization / negative regulation of protein phosphorylation / FAD binding / monooxygenase activity / actin filament / SH3 domain binding / small GTPase binding / actin filament binding / actin cytoskeleton / Factors involved in megakaryocyte development and platelet production / actin binding / midbody / endosome membrane / negative regulation of apoptotic process / protein kinase binding / signal transduction / metal ion binding / plasma membrane / cytosol / cytoplasm

ドメイン・相同性:

bMERB domain / Bivalent Mical/EHBP Rab binding domain / bMERB domain profile. / DUF3585 / : / LIM zinc-binding domain signature. / LIM domain / Zinc-binding domain present in Lin-11, Isl-1, Mec-3. / Zinc finger, LIM-type / LIM domain profile. / FAD-binding domain / FAD binding domain / Calponin homology domain / Calponin homology (CH) domain / Calponin homology domain / CH domain superfamily / Calponin homology (CH) domain profile. / FAD/NAD(P)-binding domain superfamily

構成要素:

[F-actin]-monooxygenase MICAL1

• 生物種: Homo sapiens (ヒト)
• 引用: ジャーナル: Protein Sci / 年: 2019; 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 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.

登録者

• Maxim Petoukhov (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #2178: ; タイプ: mix / ダミー原子の半径: 1.90 A / コメント: PDB file corresponds to full-length Mical1 / カイ2乗値: 1.63; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) (Truncated MOCHLIM construct); 試料濃度: 6.82 mg/ml
• 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: MoChLim buffer

要素 #1193

名称: MoChLim / タイプ: protein / 記述: [F-actin]-monooxygenase MICAL1 (MoChLim) / 分子量: 85.162 / 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 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 PTPSENSM

実験情報

• ビーム: 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.124 Å / スペクトロメータ・検出器間距離: 3 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) (Truncated MOCHLIM construct)
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 /

	Min	Max
Q	0.1436	5.0354

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 664 /

	Min	Max
Q	0.146379	1.96947
P(R) point	1	664
R	0	18

結果

D max: 18 / カーブのタイプ: sec /

	実験値	Porod
分子量	88 kDa	88 kDa
体積	-	145 nm3

	P(R)	Guinier	Guinier error
前方散乱 I0	8818	8610	15
慣性半径, Rg	4.442 nm	4 nm	0.1