+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7nwl | |||||||||||||||||||||
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タイトル | Cryo-EM structure of human integrin alpha5beta1 (open form) in complex with fibronectin and TS2/16 Fv-clasp | |||||||||||||||||||||
要素 |
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キーワード | CELL ADHESION / integrin / fibronectin / TS2/16 / plasma membrane protein / a5b1 / alpha5beta1 / focal adhesion / open conformation | |||||||||||||||||||||
機能・相同性 | 機能・相同性情報 integrin alpha6-beta1 complex / integrin alpha8-beta1 complex / integrin alpha3-beta1 complex / integrin alpha5-beta1 complex / integrin alpha7-beta1 complex / integrin alpha10-beta1 complex / integrin alpha11-beta1 complex / positive regulation of glutamate uptake involved in transmission of nerve impulse / myoblast fate specification / regulation of inward rectifier potassium channel activity ...integrin alpha6-beta1 complex / integrin alpha8-beta1 complex / integrin alpha3-beta1 complex / integrin alpha5-beta1 complex / integrin alpha7-beta1 complex / integrin alpha10-beta1 complex / integrin alpha11-beta1 complex / positive regulation of glutamate uptake involved in transmission of nerve impulse / myoblast fate specification / regulation of inward rectifier potassium channel activity / regulation of collagen catabolic process / integrin alpha9-beta1 complex / integrin alpha4-beta1 complex / cardiac cell fate specification / integrin binding involved in cell-matrix adhesion / cell-cell adhesion mediated by integrin / integrin alpha1-beta1 complex / collagen binding involved in cell-matrix adhesion / integrin alpha2-beta1 complex / Localization of the PINCH-ILK-PARVIN complex to focal adhesions / regulation of synapse pruning / reactive gliosis / formation of radial glial scaffolds / cerebellar climbing fiber to Purkinje cell synapse / Other semaphorin interactions / negative regulation of monocyte activation / Formation of the ureteric bud / positive regulation of vascular endothelial growth factor signaling pathway / calcium-independent cell-matrix adhesion / negative regulation of transforming growth factor beta production / integrin alphav-beta1 complex / CD40 signaling pathway / positive regulation of fibroblast growth factor receptor signaling pathway / Fibronectin matrix formation / Extracellular matrix organization / basement membrane organization / positive regulation of substrate-dependent cell migration, cell attachment to substrate / myelin sheath abaxonal region / neural crest cell migration involved in autonomic nervous system development / peptidase activator activity / alphav-beta3 integrin-vitronectin complex / CHL1 interactions / fibrinogen complex / Laminin interactions / cardiac muscle cell myoblast differentiation / RUNX2 regulates genes involved in cell migration / MET interacts with TNS proteins / peptide cross-linking / germ cell migration / leukocyte tethering or rolling / cardiac muscle cell differentiation / mesodermal cell differentiation / vascular endothelial growth factor receptor 2 binding / cell projection organization / Platelet Adhesion to exposed collagen / integrin activation / ALK mutants bind TKIs / myoblast fusion / Elastic fibre formation / cell-substrate junction assembly / platelet-derived growth factor receptor binding / axon extension / positive regulation of vascular endothelial growth factor receptor signaling pathway / cell migration involved in sprouting angiogenesis / positive regulation of fibroblast migration / Differentiation of keratinocytes in interfollicular epidermis in mammalian skin / wound healing, spreading of epidermal cells / myoblast differentiation / positive regulation of cell-substrate adhesion / integrin complex / heterotypic cell-cell adhesion / regulation of spontaneous synaptic transmission / biological process involved in interaction with symbiont / dendrite morphogenesis / proteoglycan binding / Basigin interactions / Molecules associated with elastic fibres / lamellipodium assembly / negative regulation of Rho protein signal transduction / heterophilic cell-cell adhesion via plasma membrane cell adhesion molecules / extracellular matrix structural constituent / cell adhesion mediated by integrin / MET activates PTK2 signaling / negative regulation of vasoconstriction / Syndecan interactions / epidermal growth factor receptor binding / leukocyte cell-cell adhesion / sarcomere organization / positive regulation of wound healing / muscle organ development / p130Cas linkage to MAPK signaling for integrins / maintenance of blood-brain barrier / positive regulation of neuroblast proliferation / cell-substrate adhesion / positive regulation of sprouting angiogenesis / endodermal cell differentiation / homophilic cell adhesion via plasma membrane adhesion molecules / TGF-beta receptor signaling activates SMADs / establishment of mitotic spindle orientation / GRB2:SOS provides linkage to MAPK signaling for Integrins 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | Homo sapiens (ヒト) Mus musculus (ハツカネズミ) | |||||||||||||||||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.1 Å | |||||||||||||||||||||
データ登録者 | Schumacher, S. / Dedden, D. / Vazquez Nunez, R. / Matoba, K. / Takagi, J. / Biertumpfel, C. / Mizuno, N. | |||||||||||||||||||||
資金援助 | ドイツ, European Union, 米国, 6件
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引用 | ジャーナル: Sci Adv / 年: 2021 タイトル: Structural insights into integrin αβ opening by fibronectin ligand. 著者: Stephanie Schumacher / Dirk Dedden / Roberto Vazquez Nunez / Kyoko Matoba / Junichi Takagi / Christian Biertümpfel / Naoko Mizuno / 要旨: Integrin αβ is a major fibronectin receptor critical for cell migration. Upon complex formation, fibronectin and αβ undergo conformational changes. While this is key for cell-tissue connections, ...Integrin αβ is a major fibronectin receptor critical for cell migration. Upon complex formation, fibronectin and αβ undergo conformational changes. While this is key for cell-tissue connections, its mechanism is unknown. Here, we report cryo-electron microscopy structures of native human αβ with fibronectin to 3.1-angstrom resolution, and in its resting state to 4.6-angstrom resolution. The αβ-fibronectin complex revealed simultaneous interactions at the arginine-glycine-aspartate loop, the synergy site, and a newly identified binding site proximal to adjacent to metal ion-dependent adhesion site, inducing the translocation of helix α1 to secure integrin opening. Resting αβ adopts an incompletely bent conformation, challenging the model of integrin sharp bending inhibiting ligand binding. Our biochemical and structural analyses showed that affinity of αβ for fibronectin is increased with manganese ions (Mn) while adopting the half-bent conformation, indicating that ligand-binding affinity does not depend on conformation, and αβ opening is induced by ligand-binding. | |||||||||||||||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 7nwl.cif.gz | 335.5 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7nwl.ent.gz | 267.5 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7nwl.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 7nwl_validation.pdf.gz | 1.4 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 7nwl_full_validation.pdf.gz | 1.5 MB | 表示 | |
XML形式データ | 7nwl_validation.xml.gz | 78 KB | 表示 | |
CIF形式データ | 7nwl_validation.cif.gz | 111.4 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/nw/7nwl ftp://data.pdbj.org/pub/pdb/validation_reports/nw/7nwl | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
-タンパク質 , 3種, 3分子 ABC
#1: タンパク質 | 分子量: 110111.555 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 組織: placenta / 参照: UniProt: P08648 |
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#2: タンパク質 | 分子量: 86338.594 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: Sequenced used from GenBank entry CAA30790 / 由来: (天然) Homo sapiens (ヒト) / 組織: placenta / 参照: UniProt: P05556 |
#3: タンパク質 | 分子量: 39981.160 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: FN1, FN / 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): gold / 参照: UniProt: P02751 |
-抗体 , 2種, 2分子 DE
#4: 抗体 | 分子量: 19463.992 Da / 分子数: 1 / 由来タイプ: 組換発現 / 詳細: Chimera: Homo sapiens, 9606 / 由来: (組換発現) Mus musculus (ハツカネズミ) / 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): gold |
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#5: 抗体 | 分子量: 18270.742 Da / 分子数: 1 / 由来タイプ: 組換発現 / 詳細: Chimera: Homo sapiens, 9660 / 由来: (組換発現) Mus musculus (ハツカネズミ) / 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): gold |
-糖 , 5種, 10分子
#6: 多糖 | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta- ...beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose #7: 多糖 | #8: 多糖 | alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2- ...alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | #9: 多糖 | 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-alpha-D-mannopyranose-(1-6)-[alpha-D-mannopyranose- ...2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-alpha-D-mannopyranose-(1-6)-[alpha-D-mannopyranose-(1-3)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | #10: 多糖 | alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1- ...alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | |
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-非ポリマー , 1種, 7分子
#11: 化合物 | ChemComp-MN / |
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-詳細
研究の焦点であるリガンドがあるか | N |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 | 名称: Ternary complex of integrin a5b1, fibronectin and TS2/16 Fv-clasp タイプ: COMPLEX / Entity ID: #1-#5 / 由来: MULTIPLE SOURCES | ||||||||||||||||||||
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分子量 | 値: .21 MDa / 実験値: NO | ||||||||||||||||||||
由来(天然) | 生物種: Homo sapiens (ヒト) | ||||||||||||||||||||
緩衝液 | pH: 7.5 | ||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 0.15 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES 詳細: Integrin a5b1 was assembled into MSPE3D1 nanodiscs containing lipids Folch fraction I lipids from bovine brain at a ratio of 1:29:3460, respectively. The assembly mix was incubated with SM-2 ...詳細: Integrin a5b1 was assembled into MSPE3D1 nanodiscs containing lipids Folch fraction I lipids from bovine brain at a ratio of 1:29:3460, respectively. The assembly mix was incubated with SM-2 BioBeads to remove DDM detergent from solubilized lipids and then purified by size-exclusion chromatography using a Superose 6 3.2/300 column and mixed with FN7-10 and TS/2/16 at stoichiometric ratios. | ||||||||||||||||||||
試料支持 | 詳細: GloQube at 20 mA / グリッドの材料: COPPER / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: Quantifoil R1.2/1.3 | ||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE-PROPANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 64000 X / 最大 デフォーカス(公称値): 3200 nm / 最小 デフォーカス(公称値): -500 nm / Cs: 2.62 mm / C2レンズ絞り径: 70 µm / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 7.39 sec. / 電子線照射量: 59.7 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 1 / 実像数: 9431 |
電子光学装置 | エネルギーフィルター名称: GIF Bioquantum / エネルギーフィルタースリット幅: 20 eV |
画像スキャン | サンプリングサイズ: 5 µm / 横: 6144 / 縦: 4096 |
-解析
ソフトウェア | 名称: PHENIX / バージョン: 1.17.1_3660: / 分類: 精密化 | ||||||||||||||||||||||||||||||||||||||||
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EMソフトウェア |
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画像処理 | 詳細: initial image processing automatically by FOCUS pipeline: gain normalization, motion correction and dose-weighting in MotionCor2 CTF estimation by GCTF | ||||||||||||||||||||||||||||||||||||||||
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 4218951 | ||||||||||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.1 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 680610 / アルゴリズム: FOURIER SPACE / クラス平均像の数: 2 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL | ||||||||||||||||||||||||||||||||||||||||
原子モデル構築 |
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精密化 | 交差検証法: NONE 立体化学のターゲット値: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||||||||||||||||||
原子変位パラメータ | Biso mean: 113.02 Å2 | ||||||||||||||||||||||||||||||||||||||||
拘束条件 |
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