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基本情報
登録情報 | ![]() | |||||||||
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タイトル | Structure of the Position 7 CMG-decorated gamma-Tubulin Ring Complex from Pig Brain | |||||||||
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![]() | Tubulin Complex / STRUCTURAL PROTEIN | |||||||||
機能・相同性 | ![]() Recruitment of mitotic centrosome proteins and complexes / negative regulation of centriole replication / regulation of mitotic cell cycle spindle assembly checkpoint / microtubule organizing center organization / polar microtubule / microtubule plus-end / gamma-tubulin complex / microtubule nucleation / microtubule bundle formation / gamma-tubulin binding ...Recruitment of mitotic centrosome proteins and complexes / negative regulation of centriole replication / regulation of mitotic cell cycle spindle assembly checkpoint / microtubule organizing center organization / polar microtubule / microtubule plus-end / gamma-tubulin complex / microtubule nucleation / microtubule bundle formation / gamma-tubulin binding / centrosome cycle / Recruitment of NuMA to mitotic centrosomes / regulation of neuron differentiation / pericentriolar material / mitotic spindle pole / centriole replication / establishment of mitotic spindle orientation / negative regulation of neuron differentiation / spindle assembly / neurogenesis / cytoplasmic microtubule organization / positive regulation of microtubule polymerization / centriole / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes / tubulin binding / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / meiotic cell cycle / chromosome segregation / brain development / microtubule cytoskeleton organization / spindle / neuron migration / spindle pole / cell junction / Regulation of PLK1 Activity at G2/M Transition / mitotic cell cycle / microtubule binding / microtubule / cytoskeleton / transcription cis-regulatory region binding / calmodulin binding / ciliary basal body / centrosome / protein-containing complex binding / protein kinase binding / positive regulation of DNA-templated transcription / perinuclear region of cytoplasm / Golgi apparatus / extracellular exosome / nucleoplasm / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.3 Å | |||||||||
![]() | Munoz-Hernandez H / Krutyholowa R / Wieczorek M | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Partial closure of the γ-tubulin ring complex by CDK5RAP2 activates microtubule nucleation. 著者: Yixin Xu / Hugo Muñoz-Hernández / Rościsław Krutyhołowa / Florina Marxer / Ferdane Cetin / Michal Wieczorek / ![]() 要旨: Microtubule nucleation is templated by the γ-tubulin ring complex (γ-TuRC), but its structure deviates from the geometry of α-/β-tubulin in the microtubule, explaining the complex's poor ...Microtubule nucleation is templated by the γ-tubulin ring complex (γ-TuRC), but its structure deviates from the geometry of α-/β-tubulin in the microtubule, explaining the complex's poor nucleating activity. Several proteins may activate the γ-TuRC, but the mechanisms underlying activation are not known. Here, we determined the structure of the porcine γ-TuRC purified using CDK5RAP2's centrosomin motif 1 (CM1). We identified an unexpected conformation of the γ-TuRC bound to multiple protein modules containing MZT2, GCP2, and CDK5RAP2, resulting in a long-range constriction of the γ-tubulin ring that brings it in closer agreement with the 13-protofilament microtubule. Additional CDK5RAP2 promoted γ-TuRC decoration and stimulated the microtubule-nucleating activities of the porcine γ-TuRC and a reconstituted, CM1-free human complex in single-molecule assays. Our results provide a structural mechanism for the control of microtubule nucleation by CM1 proteins and identify conformational transitions in the γ-TuRC that prime it for microtubule nucleation. | |||||||||
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マップデータ | ![]() | 116.2 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 21.1 KB 21.1 KB | 表示 表示 | ![]() |
画像 | ![]() | 86.7 KB | ||
Filedesc metadata | ![]() | 8.1 KB | ||
その他 | ![]() ![]() | 98.5 MB 98.5 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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マップ
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投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.41333 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: #2
ファイル | emd_51020_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #1
ファイル | emd_51020_half_map_2.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
-全体 : Gamma-Tubulin Ring Complex in native pig brain
全体 | 名称: Gamma-Tubulin Ring Complex in native pig brain |
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要素 |
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-超分子 #1: Gamma-Tubulin Ring Complex in native pig brain
超分子 | 名称: Gamma-Tubulin Ring Complex in native pig brain / タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all |
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由来(天然) | 生物種: ![]() ![]() |
-分子 #1: Gamma-tubulin complex component 3
分子 | 名称: Gamma-tubulin complex component 3 / タイプ: protein_or_peptide / ID: 1 / 詳細: SsGCP3 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 103.172477 KDa |
配列 | 文字列: MATPDQKSPN VLLQNLCCRI LGKSEADVAQ QFQYAVRVIG SNFAPTVERD EFLVAEKIKK ELTRQRREAD AALFSELHRK LHSQGVLKN KWSILYLLLS LSEDPRKQPS KVSGYAALFA QALPRDAHST PYYYARPQSL PLNYQERGAP SAQSAGSAGS S GVSSLGTY ...文字列: MATPDQKSPN VLLQNLCCRI LGKSEADVAQ QFQYAVRVIG SNFAPTVERD EFLVAEKIKK ELTRQRREAD AALFSELHRK LHSQGVLKN KWSILYLLLS LSEDPRKQPS KVSGYAALFA QALPRDAHST PYYYARPQSL PLNYQERGAP SAQSAGSAGS S GVSSLGTY ALNGPTPPPP PPALLPGQPL PAPGVGDGLR QQLGSRLAWT LTASQPSLPS TTSKAVPSSG SRGAARPRRE GD AAAGAVE VTEAALVRDI LYVFQGIDGK HVKMSNADNC YTVEGKANLS KSLRDTAVRL AELGWLHNKI RKYTDQRSLD RSF GLVGQS FCAALHQELR EYYRLLSVLH SQLQLEDDQG VNLGLESSLT LRRLLVWTYD PKMRLKTLAA LVDHCQGRKG GELA SAVHA YTKTGDPYAR SLVQHILSLV SHPVLSFLYR WIYDGELEDT YHEFFVASDP AVKADRLWHD KYALRKPMIP SFMTM DQCR KVLLIGKSIN FLHQVCHDQT PTTKMIAVTK SAESPQDAAD LFTDLENAFQ GKIDAAYFET SKYLLDVLNK KYSLLD HMQ AMRRYLLLGQ GDFIRHLMDL LKPELVRPAT TLYQHNLTGI LETAVRATNA QFDSPEILKR LDVRLLEVSP GDTGWDV FS LDYHVDGPIA TVFTRECMSH YLRAFNFLWR AKRVEYILTD IRKGHMCNAR LLRSMPEFSG VLHHCHILAS EMVHFIHQ M QYYVTFEVLE CSWDELWNRV QRAQDLDHII AAHEAFLGTV ISRCLLDSDS RALLNQLRAV FDQIIELQNT QDAIYRAAL EELQRRLQFE EKKKQREAEG QWGVSAAEEE QEKRRVQEFQ ESIPKMCSQL RILTHFYQGV VQQFLVSLTT SSDESLRFLS FRLDFNEHY RAREPRLRVS LGTRGRRSSH T UniProtKB: Gamma-tubulin complex component 3 |
-分子 #2: Gamma-tubulin complex component
分子 | 名称: Gamma-tubulin complex component / タイプ: protein_or_peptide / ID: 2 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 102.609703 KDa |
配列 | 文字列: MSEFRIHHDV NELLSLLRVH GGDGAEVYID LLQKNRTPYV TTTVSAHSAK VKIAEFSRTP EDFLKKYDEL KSKNTRNLDP LVYLLSKLM EDRETLQYLQ QNAKERAELA ASAAASSTAS FGASATASKI SMQELEELRK QLGSVATGPT WQQSLELTRK M LRDKQSKK ...文字列: MSEFRIHHDV NELLSLLRVH GGDGAEVYID LLQKNRTPYV TTTVSAHSAK VKIAEFSRTP EDFLKKYDEL KSKNTRNLDP LVYLLSKLM EDRETLQYLQ QNAKERAELA ASAAASSTAS FGASATASKI SMQELEELRK QLGSVATGPT WQQSLELTRK M LRDKQSKK NSGQRLPVLP AWVYERPALL GDFLPGTGGS ADTAVPIGSL PLASQEAAVV EDLLYVLVGV DGRYISAQPL TG RQGRTFL VDPNLDLSIR ELVSRILPVA ASYSTVTRFI EEKSSFEYGQ VNHALAAAMR TLVKEYLVLV TQLEQLQRQG LLS LQKLWF YIQPAMRSLD ILASLATSVD KGECIGGATL SLLHDRSFSY TGDSQAQELC LHLTKAASTP YFEILEKWIY RGII DDPYS EFMVEEHELR KEKIQEDYND KYWDQRYTVV QRQIPSFLQK MAGKVLSTGK YLNVVRECGH DVTCPVAKEV VYTLK ERAY VEQIEKAFSY ASKVLLDFLM GEKELLAHLR SIKRYFLMDQ GDFFVHFMDL TEEELKKPVD DITPTRLEAL LELALR MST ANTDPFKDDL KIDLMPHDLI TQLLRVLAIE TQQEKAMVHA DPTELTLSGL EAFSFDYVVT WPLSLIINRK ALTRYQM LF RHMFYCKHVE RQLCSVWISN KAAKRFSLHS AKWFAGAFTL RQRMLNFVQN IQSYMMFEVM EPTWHVLEQN LRSASNID D VLGHHASFLD NCLKDCMLTN PELLRVFSKL MSVCVMFTNC LQRFTQSMKL DSELGHPALE PGAMLGPPTE AERAEERAR KELARKCLAE HVDAPQLASS FEATITKFDK NFSAHLLDLL ARLSIYSTSD CEHGMASVIS RLDFNGFYTE RLERLSAERS QKAAPPVPG PRGPPALVPR VAVTAQ UniProtKB: Gamma-tubulin complex component |
-分子 #3: Mitotic-spindle organizing protein 2A isoform X4
分子 | 名称: Mitotic-spindle organizing protein 2A isoform X4 / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 15.920321 KDa |
配列 | 文字列: MAAPGAGPGP GAPPGLEAAL QKLALRRKKV LSAEETELFE LAQAAGGAMD PEVFKILVDL LKLNVAPLAV FQMLKSMCAG QRLASEPQD PVAVPLPTTS VPETRGRNRG SSALGGGPAL AERSGREGSS QRMPRQPSAT RLPKGGGPGK SPTRST UniProtKB: Mitotic-spindle organizing protein 2A isoform X4 |
-分子 #4: CDK5 regulatory subunit-associated protein 2
分子 | 名称: CDK5 regulatory subunit-associated protein 2 / タイプ: protein_or_peptide / ID: 4 / コピー数: 2 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 215.344219 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MMDLVLEEDV TVPGTLSGCS GLVPSVPDDL DGINPNAGLG NGLLPNVSEE TVSPTRARNM KDFENQITEL KKENFNLKLR IYFLEERMQ QEFHGPTEHI YKTNIELKVE VESLKRELQE REQLLIKASK AVESLAEAGG SEIQRVKEDA RKKVQQVEDL L TKRILLLE ...文字列: MMDLVLEEDV TVPGTLSGCS GLVPSVPDDL DGINPNAGLG NGLLPNVSEE TVSPTRARNM KDFENQITEL KKENFNLKLR IYFLEERMQ QEFHGPTEHI YKTNIELKVE VESLKRELQE REQLLIKASK AVESLAEAGG SEIQRVKEDA RKKVQQVEDL L TKRILLLE KDVTAAQAEL EKAFAGTETE KALRLRLESK LSEMKKMHEG DLAMALVLDE KDRLIEELKL SLKSKEALIQ CL KEEKSQM ACPDENVSSG ELRGLCAAPR EEKERETEAA QMEHQKERNS FEERIQALEE DLREKEREIA TEKKNSLKRD KAI QGLTMA LKSKEKKVEE LNSEIEKLSA AFAKAREALQ KAQTQEFQGS EDYETALSGK EALSAALRSQ NLTKSTENHR LRRS IKKIT QELSDLQQER ERLEKDLEEA HREKSKGDCT IRDLRNEVEK LRNEVNEREK AMENRYKSLL SESNKKLHNQ EQVIK HLTE STNQKDVLLQ KFNEKDLEVI QQNCYLMAAE DLELRSEGLI TEKCSSQQPP GSKTIFSKEK KQSSDYEELI QVLKKE QDI YTHLVKSLQE SDSINNLQAE LNKIFALRKQ LEQDVLSYQN LRKTLEEQIS EIRRREEESF SLYSDQTSYL SICLEEN NR FQVEHFSQEE LKKKVSDLIQ LVKELYTDNQ HLKKTIFDLS CMGFQGNGFP DRLASTEQTE LLASKEDEDT IKIGEDDE I NFLSDQHLQQ SNEIMKDLSK GGCKNGYLRH TESKISDCDG AHAPGCLEEG AFINLLAPLF NEKATLLLES RPDLLKVVR ELLLGQLFLT EQEVSGEHLD GKTEKTPKQK GELVHFVQTN SFSKPHDELK LSCEAQLVKA GEVPKVGLKD ASVQTVATEG DLLRFKHEA TREAWEEKPI NTALSAEHRP ENLHGVPGWQ AALLSLPGIT NREAKKSRLP ILIKPSRSLG NMYRLPATQE V VTQLQSQI LELQGELKEF KTCNKQLHQK LILAEAVMEG RPTPDKTLLN AQPPVGAAYQ DSPGEQKGIK TTSSVWRDKE MD SDQQRSY EIDSEICPPD DLASLPSCKE NPEDVLSPTS VATYLSSKSQ PSAKVSVMGT DQSESINTSN ETEYLKQKIH DLE TELEGY QNFIFQLQKH SQCSEAIITV LCGTEGAQDG LSKPKNGSDG EEMTFSSLHQ VRYVKHVKIL GPLAPEMIDS RVLE NLKQQ LEEQEYKLQK EQNLNMQLFS EIHNLQNKFR DLSPPRYDSL VQSQARELSL QRQQIKDGHG ICVISRQHMN TMIKA FEEL LQASDVDYCV AEGFQEQLNQ CAELLEKLEK LFLNGKSVGV EMNTQNELME RIEEDNLTYQ HLLPESPEPS ASHALS DYE TSEKSFFSRD QKQDNETEKT SVMVNSFSQD LLMEHIQEIR TLRKRLEESI KTNEKLRKQL ERQGSEFVQG STSIFAS GS ELHSSLTSEI HFLRKQNQAL NAMLIKGSRD KQKENDKLRE SLSRKTVSLE HLQREYASVK EENERLQKEG SEKERHNQ Q LIQEVRCSGQ ELSRVQEEVK LRQQLLSQND KLLQSLRVEL KAYEKLDEEH RRLREASGEG WKGQDPFRDL HSLLMEIQA LRLQLERSIE TSSTLQSRLK EQLARGAEKA QEGALTLAVQ AVSIPEVPLQ PDKHDGDKYP MESDNSFDLF DSSQAVTPKS VSETPPLSG NDTDSLSCDS GSSATSTPCV SRLVTGHHLW ASKNGRHVLG LIEDYEALLK QISQGQRLLA EMDIQTQEAP S STSQELGT KGPHPAPLSK FVSSVSTAKL TLEEAYRRLK LLWRVSLPED GQCPLHCEQI GEMKAEVTKL HKKLFEQEKK LQ NTMKLLQ LSKRQEKVIF DQLVVTHKIL RKARGNLELR PGGAHPGTCS PSRPGS UniProtKB: CDK5 regulatory subunit-associated protein 2 |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.5 構成要素:
詳細: Listed in "g-TuRC cryo-EM sample preparation" section. | ||||||||||||||||
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凍結 | 凍結剤: ETHANE-PROPANE / チャンバー内湿度: 100 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK III |
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電子顕微鏡法
顕微鏡 | TFS KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 55.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.7 µm / 最小 デフォーカス(公称値): 0.9 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
初期モデル | モデルのタイプ: INSILICO MODEL |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 4.3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 93207 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |