+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-7831 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | Cryo-EM structure of FLNaABD E254K bound to phalloidin-stabilized F-actin | |||||||||
マップデータ | Symmetrized map of FLNaABD-E254K bound to phalloidin-stabilized F-actin. This map has been low-pass filtered to 3.6 A and sharpened with a B-factor of -150. | |||||||||
試料 |
| |||||||||
キーワード | Actin-binding domain / Actin crosslinker / STRUCTURAL PROTEIN (タンパク質) | |||||||||
機能・相同性 | 機能・相同性情報 regulation of membrane repolarization during atrial cardiac muscle cell action potential / regulation of membrane repolarization during cardiac muscle cell action potential / establishment of Sertoli cell barrier / Myb complex / formation of radial glial scaffolds / glycoprotein Ib-IX-V complex / adenylate cyclase-inhibiting dopamine receptor signaling pathway / positive regulation of integrin-mediated signaling pathway / cytoplasmic sequestering of protein / tubulin deacetylation ...regulation of membrane repolarization during atrial cardiac muscle cell action potential / regulation of membrane repolarization during cardiac muscle cell action potential / establishment of Sertoli cell barrier / Myb complex / formation of radial glial scaffolds / glycoprotein Ib-IX-V complex / adenylate cyclase-inhibiting dopamine receptor signaling pathway / positive regulation of integrin-mediated signaling pathway / cytoplasmic sequestering of protein / tubulin deacetylation / actin crosslink formation / blood coagulation, intrinsic pathway / Striated Muscle Contraction / protein localization to bicellular tight junction / OAS antiviral response / positive regulation of actin filament bundle assembly / positive regulation of neuron migration / positive regulation of potassium ion transmembrane transport / Cell-extracellular matrix interactions / early endosome to late endosome transport / apical dendrite / cell-cell junction organization / positive regulation of neural precursor cell proliferation / positive regulation of platelet activation / protein localization to cell surface / wound healing, spreading of cells / negative regulation of transcription by RNA polymerase I / Fc-gamma receptor I complex binding / megakaryocyte development / GP1b-IX-V activation signalling / cortical cytoskeleton / receptor clustering / positive regulation of axon regeneration / actin filament bundle / SMAD binding / RHO GTPases activate PAKs / skeletal muscle thin filament assembly / striated muscle thin filament / 刷子縁 / semaphorin-plexin signaling pathway / cilium assembly / mitotic spindle assembly / potassium channel regulator activity / 上皮間葉転換 / blood vessel remodeling / skeletal muscle fiber development / axonal growth cone / stress fiber / heart morphogenesis / positive regulation of substrate adhesion-dependent cell spreading / release of sequestered calcium ion into cytosol / regulation of cell migration / protein kinase C binding / dendritic shaft / G protein-coupled receptor binding / マイクロフィラメント / protein localization to plasma membrane / synapse organization / 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与 / mRNA transcription by RNA polymerase II / establishment of protein localization / ゴルジ体 / negative regulation of DNA-binding transcription factor activity / negative regulation of protein catabolic process / cerebral cortex development / 血小板 / small GTPase binding / Z disc / kinase binding / positive regulation of protein import into nucleus / actin filament binding / cell-cell junction / マイクロフィラメント / negative regulation of neuron projection development / Platelet degranulation / GTPase binding / toxin activity / perikaryon / actin cytoskeleton organization / postsynapse / 血管新生 / positive regulation of canonical NF-kappaB signal transduction / DNA-binding transcription factor binding / transmembrane transporter binding / protein stabilization / hydrolase activity / cadherin binding / focal adhesion / glutamatergic synapse / 核小体 / negative regulation of apoptotic process / perinuclear region of cytoplasm / protein homodimerization activity / RNA binding / extracellular exosome / extracellular region / ATP binding / 生体膜 / 細胞核 / 細胞膜 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Gallus gallus (ニワトリ) / Amanita phalloides (タマゴテングタケ) | |||||||||
手法 | らせん対称体再構成法 / クライオ電子顕微鏡法 / 解像度: 3.54 Å | |||||||||
データ登録者 | Iwamoto DV / Huehn AR | |||||||||
引用 | ジャーナル: Nat Struct Mol Biol / 年: 2018 タイトル: Structural basis of the filamin A actin-binding domain interaction with F-actin. 著者: Daniel V Iwamoto / Andrew Huehn / Bertrand Simon / Clotilde Huet-Calderwood / Massimiliano Baldassarre / Charles V Sindelar / David A Calderwood / 要旨: Actin-cross-linking proteins assemble actin filaments into higher-order structures essential for orchestrating cell shape, adhesion, and motility. Missense mutations in the tandem calponin homology ...Actin-cross-linking proteins assemble actin filaments into higher-order structures essential for orchestrating cell shape, adhesion, and motility. Missense mutations in the tandem calponin homology domains of their actin-binding domains (ABDs) underlie numerous genetic diseases, but a molecular understanding of these pathologies is hampered by the lack of high-resolution structures of any actin-cross-linking protein bound to F-actin. Here, taking advantage of a high-affinity, disease-associated mutant of the human filamin A (FLNa) ABD, we combine cryo-electron microscopy and functional studies to reveal at near-atomic resolution how the first calponin homology domain (CH1) and residues immediately N-terminal to it engage actin. We further show that reorientation of CH2 relative to CH1 is required to avoid clashes with actin and to expose F-actin-binding residues on CH1. Our data explain localization of disease-associated loss-of-function mutations to FLNaCH1 and gain-of-function mutations to the regulatory FLNaCH2. Sequence conservation argues that this provides a general model for ABD-F-actin binding. | |||||||||
履歴 |
|
-構造の表示
ムービー |
ムービービューア |
---|---|
構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_7831.map.gz | 5.4 MB | EMDBマップデータ形式 | |
---|---|---|---|---|
ヘッダ (付随情報) | emd-7831-v30.xml emd-7831.xml | 22.3 KB 22.3 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_7831_fsc.xml | 7.4 KB | 表示 | FSCデータファイル |
画像 | emd_7831.png | 74 KB | ||
マスクデータ | emd_7831_msk_1.map | 35.3 MB | マスクマップ | |
Filedesc metadata | emd-7831.cif.gz | 6.9 KB | ||
その他 | emd_7831_additional.map.gz emd_7831_half_map_1.map.gz emd_7831_half_map_2.map.gz | 7.4 MB 11.9 MB 11.9 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-7831 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-7831 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
---|---|
「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_7831.map.gz / 形式: CCP4 / 大きさ: 35.3 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Symmetrized map of FLNaABD-E254K bound to phalloidin-stabilized F-actin. This map has been low-pass filtered to 3.6 A and sharpened with a B-factor of -150. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.33 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
|
-添付データ
-マスク #1
ファイル | emd_7831_msk_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-追加マップ: Symmetrized map of FLNaABD-E254K bound to phalloidin-stabilized F-actin....
ファイル | emd_7831_additional.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Symmetrized map of FLNaABD-E254K bound to phalloidin-stabilized F-actin. Low-pass filtered to 10 A and sharpened with a B-factor of -150. Masked to display the FLNaABD CH2 domain cryo-EM density. | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: Independently refined half map(2) of FLNaABD-E254K bound to...
ファイル | emd_7831_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Independently refined half map(2) of FLNaABD-E254K bound to phalloidin-stabilized F-actin. This half map was symmetrized using parameters calculated from the full map. | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: Independently refined half map(1) of FLNaABD-E254K bound to...
ファイル | emd_7831_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Independently refined half map(1) of FLNaABD-E254K bound to phalloidin-stabilized F-actin. This half map was symmetrized using parameters calculated from the full map. | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-試料の構成要素
-全体 : Helical complex of FLNaABD E254K bound to phalloidin-stabilized F...
全体 | 名称: Helical complex of FLNaABD E254K bound to phalloidin-stabilized F-actin |
---|---|
要素 |
|
-超分子 #1: Helical complex of FLNaABD E254K bound to phalloidin-stabilized F...
超分子 | 名称: Helical complex of FLNaABD E254K bound to phalloidin-stabilized F-actin タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#3 |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
-分子 #1: Filamin-A
分子 | 名称: Filamin-A / タイプ: protein_or_peptide / ID: 1 / コピー数: 5 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 34.47625 KDa |
組換発現 | 生物種: Escherichia coli (大腸菌) |
配列 | 文字列: MHHHHHHGSL VPRSENLYFQ GSDILQGTMS SSHSRAGQSA AGAAPGGGVD TRDAEMPATE KDLAEDAPWK KIQQNTFTRW CNEHLKCVS KRIANLQTDL SDGLRLIALL EVLSQKKMHR KHNQRPTFRQ MQLENVSVAL EFLDRESIKL VSIDSKAIVD G NLKLILGL ...文字列: MHHHHHHGSL VPRSENLYFQ GSDILQGTMS SSHSRAGQSA AGAAPGGGVD TRDAEMPATE KDLAEDAPWK KIQQNTFTRW CNEHLKCVS KRIANLQTDL SDGLRLIALL EVLSQKKMHR KHNQRPTFRQ MQLENVSVAL EFLDRESIKL VSIDSKAIVD G NLKLILGL IWTLILHYSI SMPMWDEEED EEAKKQTPKQ RLLGWIQNKL PQLPITNFSR DWQSGRALGA LVDSCAPGLC PD WDSWDAS KPVTNAREAM QQADDWLGIP QVITPEEIVD PNVDKHSVMT YLSQFPKAKL KPGAPLRPK UniProtKB: Filamin-A |
-分子 #2: Actin, alpha skeletal muscle
分子 | 名称: Actin, alpha skeletal muscle / タイプ: protein_or_peptide / ID: 2 / コピー数: 5 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Gallus gallus (ニワトリ) / 組織: Breast |
分子量 | 理論値: 41.862613 KDa |
配列 | 文字列: DEDETTALVC DNGSGLVKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIITNWD DMEKIWHHT FYNELRVAPE EHPTLLTEAP LNPKANREKM TQIMFETFNV PAMYVAIQAV LSLYASGRTT GIVLDSGDGV T HNVPIYEG ...文字列: DEDETTALVC DNGSGLVKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIITNWD DMEKIWHHT FYNELRVAPE EHPTLLTEAP LNPKANREKM TQIMFETFNV PAMYVAIQAV LSLYASGRTT GIVLDSGDGV T HNVPIYEG YALPHAIMRL DLAGRDLTDY LMKILTERGY SFVTTAEREI VRDIKEKLCY VALDFENEMA TAASSSSLEK SY ELPDGQV ITIGNERFRC PETLFQPSFI GMESAGIHET TYNSIMKCDI DIRKDLYANN VMSGGTTMYP GIADRMQKEI TAL APSTMK IKIIAPPERK YSVWIGGSIL ASLSTFQQMW ITKQEYDEAG PSIVHRKCF UniProtKB: Actin, alpha skeletal muscle |
-分子 #3: Phalloidin
分子 | 名称: Phalloidin / タイプ: protein_or_peptide / ID: 3 / コピー数: 3 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Amanita phalloides (タマゴテングタケ) |
分子量 | 理論値: 808.899 Da |
配列 | 文字列: (HYP)AW(G5G)A(ALO)C |
-分子 #4: ADENOSINE-5'-DIPHOSPHATE
分子 | 名称: ADENOSINE-5'-DIPHOSPHATE / タイプ: ligand / ID: 4 / コピー数: 5 / 式: ADP |
---|---|
分子量 | 理論値: 427.201 Da |
Chemical component information | ChemComp-ADP: |
-分子 #5: MAGNESIUM ION
分子 | 名称: MAGNESIUM ION / タイプ: ligand / ID: 5 / コピー数: 5 / 式: MG |
---|---|
分子量 | 理論値: 24.305 Da |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
解析 | らせん対称体再構成法 |
試料の集合状態 | filament |
-試料調製
緩衝液 | pH: 8 |
---|---|
凍結 | 凍結剤: ETHANE / 装置: HOMEMADE PLUNGER |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
---|---|
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 50.0 µm / 倍率(補正後): 37500 / 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / 最大 デフォーカス(公称値): 2.9 µm / 最小 デフォーカス(公称値): 1.0 µm / 倍率(公称値): 105000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / デジタル化 - 画像ごとのフレーム数: 1-40 / 撮影したグリッド数: 2 / 実像数: 2140 / 平均露光時間: 12.0 sec. / 平均電子線量: 50.0 e/Å2 詳細: Micrographs from only one of the grids were used in the reconstruction. From that grid, only micrographs where Gctf detected signal at resolutions better than 4A were used in the reconstruction (~15%). |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
-原子モデル構築 1
初期モデル |
| ||||||
---|---|---|---|---|---|---|---|
精密化 | プロトコル: RIGID BODY FIT | ||||||
得られたモデル | PDB-6d8c: |