+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-26213 | |||||||||
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タイトル | the apo structure of human mTORC2 complex | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | complex / SIGNALING PROTEIN | |||||||||
機能・相同性 | 機能・相同性情報 TORC2 signaling / regulation of peptidyl-serine phosphorylation / RNA polymerase III type 2 promoter sequence-specific DNA binding / RNA polymerase III type 1 promoter sequence-specific DNA binding / positive regulation of cytoplasmic translational initiation / T-helper 1 cell lineage commitment / positive regulation of pentose-phosphate shunt / regulation of locomotor rhythm / positive regulation of wound healing, spreading of epidermal cells / TORC2 complex ...TORC2 signaling / regulation of peptidyl-serine phosphorylation / RNA polymerase III type 2 promoter sequence-specific DNA binding / RNA polymerase III type 1 promoter sequence-specific DNA binding / positive regulation of cytoplasmic translational initiation / T-helper 1 cell lineage commitment / positive regulation of pentose-phosphate shunt / regulation of locomotor rhythm / positive regulation of wound healing, spreading of epidermal cells / TORC2 complex / cellular response to leucine starvation / TFIIIC-class transcription factor complex binding / regulation of membrane permeability / heart valve morphogenesis / negative regulation of lysosome organization / nucleus localization / RNA polymerase III type 3 promoter sequence-specific DNA binding / TORC1 complex / positive regulation of transcription of nucleolar large rRNA by RNA polymerase I / regulation of cellular response to oxidative stress / calcineurin-NFAT signaling cascade / regulation of osteoclast differentiation / voluntary musculoskeletal movement / TORC1 signaling / positive regulation of keratinocyte migration / phosphatidic acid binding / cellular response to L-leucine / Amino acids regulate mTORC1 / MTOR signalling / cellular response to nutrient / cellular response to methionine / Energy dependent regulation of mTOR by LKB1-AMPK / regulation of autophagosome assembly / energy reserve metabolic process / negative regulation of cell size / ruffle organization / negative regulation of Ras protein signal transduction / phosphatidylinositol-3,4-bisphosphate binding / cellular response to osmotic stress / phosphatidylinositol-3,5-bisphosphate binding / anoikis / cardiac muscle cell development / negative regulation of protein localization to nucleus / embryo development ending in birth or egg hatching / regulation of establishment of cell polarity / positive regulation of transcription by RNA polymerase III / negative regulation of calcineurin-NFAT signaling cascade / regulation of myelination / positive regulation of actin filament polymerization / regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / regulation of cell size / negative regulation of macroautophagy / positive regulation of oligodendrocyte differentiation / lysosome organization / Macroautophagy / positive regulation of myotube differentiation / behavioral response to pain / oligodendrocyte differentiation / Constitutive Signaling by AKT1 E17K in Cancer / mTORC1-mediated signalling / germ cell development / phosphatidylinositol-3,4,5-trisphosphate binding / CD28 dependent PI3K/Akt signaling / : / HSF1-dependent transactivation / positive regulation of TOR signaling / neuronal action potential / response to amino acid / TOR signaling / 'de novo' pyrimidine nucleobase biosynthetic process / endomembrane system / regulation of macroautophagy / positive regulation of translational initiation / cellular response to nutrient levels / positive regulation of lamellipodium assembly / phosphorylation / phagocytic vesicle / positive regulation of lipid biosynthetic process / heart morphogenesis / positive regulation of epithelial to mesenchymal transition / cardiac muscle contraction / regulation of cellular response to heat / phosphatidylinositol-4,5-bisphosphate binding / positive regulation of stress fiber assembly / cytoskeleton organization / positive regulation of endothelial cell proliferation / T cell costimulation / substantia nigra development / cellular response to starvation / cellular response to amino acid starvation / post-embryonic development / positive regulation of glycolytic process / protein serine/threonine kinase activator activity / negative regulation of autophagy / response to nutrient / response to nutrient levels / VEGFR2 mediated vascular permeability / regulation of signal transduction by p53 class mediator / Regulation of PTEN gene transcription / positive regulation of translation 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.28 Å | |||||||||
データ登録者 | Yu Z / Chen J | |||||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: J Biol Chem / 年: 2022 タイトル: Interactions between mTORC2 core subunits Rictor and mSin1 dictate selective and context-dependent phosphorylation of substrate kinases SGK1 and Akt. 著者: Zanlin Yu / Junliang Chen / Enzo Takagi / Feng Wang / Bidisha Saha / Xi Liu / Lydia-Marie Joubert / Catherine E Gleason / Mingliang Jin / Chengmin Li / Carlos Nowotny / David Agard / Yifan Cheng / David Pearce / 要旨: Mechanistic target of rapamycin complex 2 (mTORC2) is a multi-subunit kinase complex, central to multiple essential signaling pathways. Two core subunits, Rictor and mSin1, distinguish it from the ...Mechanistic target of rapamycin complex 2 (mTORC2) is a multi-subunit kinase complex, central to multiple essential signaling pathways. Two core subunits, Rictor and mSin1, distinguish it from the related mTORC1 and support context-dependent phosphorylation of its substrates. mTORC2 structures have been determined previously; however, important questions remain, particularly regarding the structural determinants mediating substrate specificity and context-dependent activity. Here, we used cryo-EM to obtain high-resolution structures of the human mTORC2 apo-complex in the presence of substrates Akt and SGK1. Using functional assays, we then tested predictions suggested by substrate-induced structural changes in mTORC2. For the first time, we visualized in the apo-state the side chain interactions between Rictor and mTOR that sterically occlude recruitment of mTORC1 substrates and confer resistance to the mTORC1 inhibitor rapamycin. Also in the apo-state, we observed that mSin1 formed extensive contacts with Rictor via a pair of short α-helices nestled between two Rictor helical repeat clusters, as well as by an extended strand that makes multiple weak contacts with Rictor helical cluster 1. In co-complex structures, we found that SGK1, but not Akt, markedly altered the conformation of the mSin1 N-terminal extended strand, disrupting multiple weak interactions while inducing a large rotation of mSin1 residue Arg-83, which then interacts with a patch of negatively charged residues within Rictor. Finally, we demonstrate mutation of Arg-83 to Ala selectively disrupts mTORC2-dependent phosphorylation of SGK1, but not of Akt, supporting context-dependent substrate selection. These findings provide new structural and functional insights into mTORC2 specificity and context-dependent activity. | |||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_26213.map.gz | 109.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-26213-v30.xml emd-26213.xml | 18.4 KB 18.4 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_26213.png | 27 KB | ||
Filedesc metadata | emd-26213.cif.gz | 8.8 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-26213 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-26213 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_26213_validation.pdf.gz | 663.4 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_26213_full_validation.pdf.gz | 663 KB | 表示 | |
XML形式データ | emd_26213_validation.xml.gz | 7 KB | 表示 | |
CIF形式データ | emd_26213_validation.cif.gz | 8 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26213 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26213 | HTTPS FTP |
-関連構造データ
関連構造データ | 7tzoMC M: このマップから作成された原子モデル C: 同じ文献を引用 (文献) |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_26213.map.gz / 形式: CCP4 / 大きさ: 129.7 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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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: 1.1 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
-試料の構成要素
-全体 : apostate of mTORC2 complex, composed of mTOR, Rictor, mLST8 and mSin1
全体 | 名称: apostate of mTORC2 complex, composed of mTOR, Rictor, mLST8 and mSin1 |
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要素 |
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-超分子 #1: apostate of mTORC2 complex, composed of mTOR, Rictor, mLST8 and mSin1
超分子 | 名称: apostate of mTORC2 complex, composed of mTOR, Rictor, mLST8 and mSin1 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: Homo sapiens (ヒト) |
-分子 #1: Serine/threonine-protein kinase mTOR
分子 | 名称: Serine/threonine-protein kinase mTOR / タイプ: protein_or_peptide / ID: 1 / コピー数: 2 / 光学異性体: LEVO / EC番号: non-specific serine/threonine protein kinase |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 302.330406 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: MVTTLSGLSG EQGPSGDMTT EEDSATHIKF SKRDEDGREL AGATMELRDS SGKTISTWIS DGHVKDFYLY PGKYTFVETA APDGYEVAT PIEFTVNEDG QVTVDGEATE GDAHTGSSGS GSGTGSMLGT GPAAATTAAT TSSNVSVLQQ FASGLKSRNE E TRAKAAKE ...文字列: MVTTLSGLSG EQGPSGDMTT EEDSATHIKF SKRDEDGREL AGATMELRDS SGKTISTWIS DGHVKDFYLY PGKYTFVETA APDGYEVAT PIEFTVNEDG QVTVDGEATE GDAHTGSSGS GSGTGSMLGT GPAAATTAAT TSSNVSVLQQ FASGLKSRNE E TRAKAAKE LQHYVTMELR EMSQEESTRF YDQLNHHIFE LVSSSDANER KGGILAIASL IGVEGGNATR IGRFANYLRN LL PSNDPVV MEMASKAIGR LAMAGDTFTA EYVEFEVKRA LEWLGADRNE GRRHAAVLVL RELAISVPTF FFQQVQPFFD NIF VAVWDP KQAIREGAVA ALRACLILTT QREPKEMQKP QWYRHTFEEA EKGFDETLAK EKGMNRDDRI HGALLILNEL VRIS SMEGE RLREEMEEIT QQQLVHDKYC KDLMGFGTKP RHITPFTSFQ AVQPQQSNAL VGLLGYSSHQ GLMGFGTSPS PAKST LVES RCCRDLMEEK FDQVCQWVLK CRNSKNSLIQ MTILNLLPRL AAFRPSAFTD TQYLQDTMNH VLSCVKKEKE RTAAFQ ALG LLSVAVRSEF KVYLPRVLDI IRAALPPKDF AHKRQKAMQV DATVFTCISM LARAMGPGIQ QDIKELLEPM LAVGLSP AL TAVLYDLSRQ IPQLKKDIQD GLLKMLSLVL MHKPLRHPGM PKGLAHQLAS PGLTTLPEAS DVGSITLALR TLGSFEFE G HSLTQFVRHC ADHFLNSEHK EIRMEAARTC SRLLTPSIHL ISGHAHVVSQ TAVQVVADVL SKLLVVGITD PDPDIRYCV LASLDERFDA HLAQAENLQA LFVALNDQVF EIRELAICTV GRLSSMNPAF VMPFLRKMLI QILTELEHSG IGRIKEQSAR MLGHLVSNA PRLIRPYMEP ILKALILKLK DPDPDPNPGV INNVLATIGE LAQVSGLEMR KWVDELFIII MDMLQDSSLL A KRQVALWT LGQLVASTGY VVEPYRKYPT LLEVLLNFLK TEQNQGTRRE AIRVLGLLGA LDPYKHKVNI GMIDQSRDAS AV SLSESKS SQDSSDYSTS EMLVNMGNLP LDEFYPAVSM VALMRIFRDQ SLSHHHTMVV QAITFIFKSL GLKCVQFLPQ VMP TFLNVI RVCDGAIREF LFQQLGMLVS FVKSHIRPYM DEIVTLMREF WVMNTSIQST IILLIEQIVV ALGGEFKLYL PQLI PHMLR VFMHDNSPGR IVSIKLLAAI QLFGANLDDY LHLLLPPIVK LFDAPEAPLP SRKAALETVD RLTESLDFTD YASRI IHPI VRTLDQSPEL RSTAMDTLSS LVFQLGKKYQ IFIPMVNKVL VRHRINHQRY DVLICRIVKG YTLADEEEDP LIYQHR MLR SGQGDALASG PVETGPMKKL HVSTINLQKA WGAARRVSKD DWLEWLRRLS LELLKDSSSP SLRSCWALAQ AYNPMAR DL FNAAFVSCWS ELNEDQQDEL IRSIELALTS QDIAEVTQTL LNLAEFMEHS DKGPLPLRDD NGIVLLGERA AKCRAYAK A LHYKELEFQK GPTPAILESL ISINNKLQQP EAAAGVLEYA MKHFGELEIQ ATWYEKLHEW EDALVAYDKK MDTNKDDPE LMLGRMRCLE ALGEWGQLHQ QCCEKWTLVN DETQAKMARM AAAAAWGLGQ WDSMEEYTCM IPRDTHDGAF YRAVLALHQD LFSLAQQCI DKARDLLDAE LTAMAGESYS RAYGAMVSCH MLSELEEVIQ YKLVPERREI IRQIWWERLQ GCQRIVEDWQ K ILMVRSLV VSPHEDMRTW LKYASLCGKS GRLALAHKTL VLLLGVDPSR QLDHPLPTVH PQVTYAYMKN MWKSARKIDA FQ HMQHFVQ TMQQQAQHAI ATEDQQHKQE LHKLMARCFL KLGEWQLNLQ GINESTIPKV LQYYSAATEH DRSWYKAWHA WAV MNFEAV LHYKHQNQAR DEKKKLRHAS GANITNATTA ATTAATATTT ASTEGSNSES EAESTENSPT PSPLQKKVTE DLSK TLLMY TVPAVQGFFR SISLSRGNNL QDTLRVLTLW FDYGHWPDVN EALVEGVKAI QIDTWLQVIP QLIARIDTPR PLVGR LIHQ LLTDIGRYHP QALIYPLTVA SKSTTTARHN AANKILKNMC EHSNTLVQQA MMVSEELIRV AILWHEMWHE GLEEAS RLY FGERNVKGMF EVLEPLHAMM ERGPQTLKET SFNQAYGRDL MEAQEWCRKY MKSGNVKDLT QAWDLYYHVF RRISKQL PQ LTSLELQYVS PKLLMCRDLE LAVPGTYDPN QPIIRIQSIA PSLQVITSKQ RPRKLTLMGS NGHEFVFLLK GHEDLRQD E RVMQLFGLVN TLLANDPTSL RKNLSIQRYA VIPLSTNSGL IGWVPHCDTL HALIRDYREK KKILLNIEHR IMLRMAPDY DHLTLMQKVE VFEHAVNNTA GDDLAKLLWL KSPSSEVWFD RRTNYTRSLA VMSMVGYILG LGDRHPSNLM LDRLSGKILH IDFGDCFEV AMTREKFPEK IPFRLTRMLT NAMEVTGLDG NYRITCHTVM EVLREHKDSV MAVLEAFVYD PLLNWRLMDT N TKGNKRSR TRTDSYSAGQ SVEILDGVEL GEPAHKKTGT TVPESIHSFI GDGLVKPEAL NKKAIQIINR VRDKLTGRDF SH DDTLDVP TQVELLIKQA TSHENLCQCY IGWCPFW UniProtKB: Serine/threonine-protein kinase mTOR |
-分子 #2: Target of rapamycin complex subunit LST8
分子 | 名称: Target of rapamycin complex subunit LST8 / タイプ: protein_or_peptide / ID: 2 / コピー数: 2 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 37.998254 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: MGYPYDVPDY ADLNGGGGGS TMNTSPGTVG SDPVILATAG YDHTVRFWQA HSGICTRTVQ HQDSQVNALE VTPDRSMIAA AGYQHIRMY DLNSNNPNPI ISYDGVNKNI ASVGFHEDGR WMYTGGEDCT ARIWDLRSRN LQCQRIFQVN APINCVCLHP N QAELIVGD ...文字列: MGYPYDVPDY ADLNGGGGGS TMNTSPGTVG SDPVILATAG YDHTVRFWQA HSGICTRTVQ HQDSQVNALE VTPDRSMIAA AGYQHIRMY DLNSNNPNPI ISYDGVNKNI ASVGFHEDGR WMYTGGEDCT ARIWDLRSRN LQCQRIFQVN APINCVCLHP N QAELIVGD QSGAIHIWDL KTDHNEQLIP EPEVSITSAH IDPDASYMAA VNSTGNCYVW NLTGGIGDEV TQLIPKTKIP AH TRYALQC RFSPDSTLLA TCSADQTCKI WRTSNFSLMT ELSIKSGNPG ESSRGWMWGC AFSGDSQYIV TASSDNLARL WCV ETGEIK REYGGHQKAV VCLAFNDSVL G UniProtKB: Target of rapamycin complex subunit LST8 |
-分子 #3: Rapamycin-insensitive companion of mTOR
分子 | 名称: Rapamycin-insensitive companion of mTOR / タイプ: protein_or_peptide / ID: 3 / コピー数: 2 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 193.846328 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: MDYKDDDDKG STMAAIGRGR SLKNLRVRGR NDSGEENVPL DLTREPSDNL REILQNVARL QGVSNMRKLG HLNNFTKLLC DIGHSEEKL GFHYEDIIIC LRLALLNEAK EVRAAGLRAL RYLIQDSSIL QKVLKLKVDY LIARCIDIQQ SNEVERTQAL R LVRKMITV ...文字列: MDYKDDDDKG STMAAIGRGR SLKNLRVRGR NDSGEENVPL DLTREPSDNL REILQNVARL QGVSNMRKLG HLNNFTKLLC DIGHSEEKL GFHYEDIIIC LRLALLNEAK EVRAAGLRAL RYLIQDSSIL QKVLKLKVDY LIARCIDIQQ SNEVERTQAL R LVRKMITV NASLFPSSVT NSLIAVGNDG LQERDRMVRA CIAIICELAL QNPEVVALRG GLNTILKNVI DCQLSRINEA LI TTILHLL NHPKTRQYVR ADVELERILA PYTDFHYRHS PDTAEGQLKE DREARFLASK MGIIATFRSW AGIINLCKPG NSG IQSLIG VLCIPNMEIR RGLLEVLYDI FRLPLPVVTE EFIEALLSVD PGRFQDSWRL SDGFVAAEAK TILPHRARSR PDLM DNYLA LILSAFIRNG LLEGLVEVIT NSDDHISVRA TILLGELLHM ANTILPHSHS HHLHCLPTLM NMAASFDIPK EKRLR ASAA LNCLKRFHEM KKRGPKPYSL HLDHIIQKAI ATHQKRDQYL RVQKDIFILK DTEEALLINL RDSQVLQHKE NLEWNW NLI GTILKWPNVN LRNYKDEQLH RFVRRLLYFY KPSSKLYANL DLDFAKAKQL TVVGCQFTEF LLESEEDGQG YLEDLVK DI VQWLNASSGM KPERSLQNNG LLTTLSQHYF LFIGTLSCHP HGVKMLEKCS VFQCLLNLCS LKNQDHLLKL TVSSLDYS R DGLARVILSK ILTAATDACR LYATKHLRVL LRANVEFFNN WGIELLVTQL HDKNKTISSE ALDILDEACE DKANLHALI QMKPALSHLG DKGLLLLLRF LSIPKGFSYL NERGYVAKQL EKWHREYNSK YVDLIEEQLN EALTTYRKPV DGDNYVRRSN QRLQRPHVY LPIHLYGQLV HHKTGCHLLE VQNIITELCR NVRTPDLDKW EEIKKLKASL WALGNIGSSN WGLNLLQEEN V IPDILKLA KQCEVLSIRG TCVYVLGLIA KTKQGCDILK CHNWDAVRHS RKHLWPVVPD DVEQLCNELS SIPSTLSLNS ES TSSRHNS ESESVPSSMF ILEDDRFGSS STSTFFLDIN EDTEPTFYDR SGPIKDKNSF PFFASSKLVK NRILNSLTLP NKK HRSSSD PKGGKLSSES KTSNRRIRTL TEPSVDFNHS DDFTPISTVQ KTLQLETSFM GNKHIEDTGS TPSIGENDLK FTKN FGTEN HRENTSRERL VVESSTSSHM KIRSQSFNTD TTTSGISSMS SSPSRETVGV DATTMDTDCG SMSTVVSTKT IKTSH YLTP QSNHLSLSKS NSVSLVPPGS SHTLPRRAQS LKAPSIATIK SLADCNFSYT SSRDAFGYAT LKRLQQQRMH PSLSHS EAL ASPAKDVLFT DTITMKANSF ESRLTPSRFM KALSYASLDK EDLLSPINQN TLQRSSSVRS MVSSATYGGS DDYIGLA LP VDINDIFQVK DIPYFQTKNI PPHDDRGARA FAHDAGGLPS GTGGLVKNSF HLLRQQMSLT EIMNSIHSDA SLFLESTE D TGLQEHTDDN CLYCVCIEIL GFQPSNQLSA ICSHSDFQDI PYSDWCEQTI HNPLEVVPSK FSGISGCSDG VSQEGSASS TKSTELLLGV KTIPDDTPMC RILLRKEVLR LVINLSSSVS TKCHETGLLT IKEKYPQTFD DICLYSEVSH LLSHCTFRLP CRRFIQELF QDVQFLQMHE EAEAVLATPP KQPIVDTSAE S UniProtKB: Rapamycin-insensitive companion of mTOR |
-分子 #4: Target of rapamycin complex 2 subunit MAPKAP1
分子 | 名称: Target of rapamycin complex 2 subunit MAPKAP1 / 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: Homo sapiens (ヒト) |
分子量 | 理論値: 60.732328 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: MAFLDNPTII LAHIRQSHVT SDDTGMCEMV LIDHDVDLEK IHPPSMPGDS GSEIQGSNGE TQGYVYAQSV DITSSWDFGI RRRSNTAQR LERLRKERQN QIKCKNIQWK ERNSKQSAQE LKSLFEKKSL KEKPPISGKQ SILSVRLEQC PLQLNNPFNE Y SKFDGKGH ...文字列: MAFLDNPTII LAHIRQSHVT SDDTGMCEMV LIDHDVDLEK IHPPSMPGDS GSEIQGSNGE TQGYVYAQSV DITSSWDFGI RRRSNTAQR LERLRKERQN QIKCKNIQWK ERNSKQSAQE LKSLFEKKSL KEKPPISGKQ SILSVRLEQC PLQLNNPFNE Y SKFDGKGH VGTTATKKID VYLPLHSSQD RLLPMTVVTM ASARVQDLIG LICWQYTSEG REPKLNDNVS AYCLHIAEDD GE VDTDFPP LDSNEPIHKF GFSTLALVEK YSSPGLTSKE SLFVRINAAH GFSLIQVDNT KVTMKEILLK AVKRRKGSQK VSG PQYRLE KQSEPNVAVD LDSTLESQSA WEFCLVRENS SRADGVFEED SQIDIATVQD MLSSHHYKSF KVSMIHRLRF TTDV QLGIS GDKVEIDPVT NQKASTKFWI KQKPISIDSD LLCACDLAEE KSPSHAIFKL TYLSNHDYKH LYFESDAATV NEIVL KVNY ILESRASTAR ADYFAQKQRK LNRRTSFSFQ KEKKSGQQAA AGGGGYPYDV PDYA UniProtKB: Target of rapamycin complex 2 subunit MAPKAP1 |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | 3D array |
-試料調製
濃度 | 1 mg/mL |
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緩衝液 | pH: 7.5 |
グリッド | モデル: Homemade / 材質: GOLD / メッシュ: 300 / 支持フィルム - 材質: GRAPHENE OXIDE / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 0.2 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 22 K / 装置: FEI VITROBOT MARK IV |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 60.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.0 µm / 最小 デフォーカス(公称値): 1.0 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
初期モデル | モデルのタイプ: NONE |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 3.28 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 288538 |
初期 角度割当 | タイプ: NOT APPLICABLE |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |