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データを開く
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基本情報
| 登録情報 | ![]() | |||||||||
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| タイトル | AP-3 Arf1 dimeric interface, focused refinement | |||||||||
マップデータ | deepEMHancer sharpened map for Arf1 dimer | |||||||||
試料 |
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キーワード | Adaptor Protein complex / Endosomal Trafficking / Lysosomal Trafficking / Protein transport / AP complex / TRANSPORT PROTEIN | |||||||||
| 機能・相同性 | 機能・相同性情報: / clathrin-coated vesicle cargo loading, AP-3-mediated / skin epidermis development / regulation of organelle transport along microtubule / AP-3 adaptor complex / anterograde synaptic vesicle transport / granzyme-mediated programmed cell death signaling pathway / phagolysosome membrane / microvesicle / positive regulation of natural killer cell degranulation ...: / clathrin-coated vesicle cargo loading, AP-3-mediated / skin epidermis development / regulation of organelle transport along microtubule / AP-3 adaptor complex / anterograde synaptic vesicle transport / granzyme-mediated programmed cell death signaling pathway / phagolysosome membrane / microvesicle / positive regulation of natural killer cell degranulation / cytolytic granule membrane / establishment of protein localization to organelle / postsynaptic recycling endosome / mitotic cleavage furrow ingression / trans-Golgi Network Vesicle Budding / platelet dense granule organization / melanosome assembly / Glycosphingolipid transport / clathrin adaptor complex / regulation of receptor internalization / Golgi to lysosome transport / Intra-Golgi traffic / granulocyte differentiation / regulation of Arp2/3 complex-mediated actin nucleation / positive regulation of NK T cell differentiation / postsynaptic neurotransmitter receptor internalization / Synthesis of PIPs at the Golgi membrane / clathrin-coated vesicle membrane / lysosomal lumen acidification / antigen processing and presentation, exogenous lipid antigen via MHC class Ib / respiratory system process / protein targeting to lysosome / melanosome organization / GTP-dependent protein binding / anterograde axonal transport / Nef Mediated CD4 Down-regulation / intracellular zinc ion homeostasis / positive regulation of natural killer cell mediated cytotoxicity / dendritic spine organization / protein localization to cell surface / long-term synaptic depression / COPI-dependent Golgi-to-ER retrograde traffic / azurophil granule membrane / toll-like receptor signaling pathway / Lysosome Vesicle Biogenesis / lysosome organization / Golgi Associated Vesicle Biogenesis / ion channel inhibitor activity / Synthesis of PIPs at the plasma membrane / Association of TriC/CCT with target proteins during biosynthesis / cell leading edge / autolysosome / lung morphogenesis / autophagosome membrane / homeostasis of number of cells / ficolin-1-rich granule membrane / intracellular copper ion homeostasis / single fertilization / hematopoietic progenitor cell differentiation / COPI-mediated anterograde transport / vesicle-mediated transport / multivesicular body / axon cytoplasm / MHC class II antigen presentation / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / cytoplasmic vesicle membrane / small monomeric GTPase / sarcomere / intracellular protein transport / cellular response to virus / sarcolemma / mRNA transcription by RNA polymerase II / small GTPase binding / cell morphogenesis / endocytosis / blood coagulation / synaptic vesicle / Signaling by BRAF and RAF1 fusions / melanosome / late endosome / late endosome membrane / virus receptor activity / G protein activity / cytoplasmic vesicle / protein phosphatase binding / spermatogenesis / early endosome / lysosome / endosome membrane / neuron projection / postsynaptic density / protein stabilization / inflammatory response / Golgi membrane / external side of plasma membrane / protein domain specific binding / lysosomal membrane / focal adhesion / GTPase activity / Neutrophil degranulation 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: 4.2 Å | |||||||||
データ登録者 | Begley MC / Baker RW | |||||||||
| 資金援助 | 米国, 1件
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引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2024タイトル: A structure-based mechanism for initiation of AP-3 coated vesicle formation. 著者: Matthew Begley / Mahira Aragon / Richard W Baker / ![]() 要旨: Adaptor protein complex-3 (AP-3) mediates cargo sorting from endosomes to lysosomes and lysosome-related organelles. Recently, it was shown that AP-3 adopts a constitutively open conformation ...Adaptor protein complex-3 (AP-3) mediates cargo sorting from endosomes to lysosomes and lysosome-related organelles. Recently, it was shown that AP-3 adopts a constitutively open conformation compared to the related AP-1 and AP-2 coat complexes, which are inactive until undergoing large conformational changes upon membrane recruitment. How AP-3 is regulated is therefore an open question. To understand the mechanism of AP-3 membrane recruitment and activation, we reconstituted human AP-3 and determined multiple structures in the soluble and membrane-bound states using electron cryo-microscopy. Similar to yeast AP-3, human AP-3 is in a constitutively open conformation. To reconstitute AP-3 activation by adenosine di-phosphate (ADP)-ribosylation factor 1 (Arf1), a small guanosine tri-phosphate (GTP)ase, we used lipid nanodiscs to build Arf1-AP-3 complexes on membranes and determined three structures showing the stepwise conformational changes required for formation of AP-3 coated vesicles. First, membrane recruitment is driven by one of two predicted Arf1 binding sites, which flexibly tethers AP-3 to the membrane. Second, cargo binding causes AP-3 to adopt a fixed position and rigidifies the complex, which stabilizes binding for a second Arf1 molecule. Finally, binding of the second Arf1 molecule provides the template for AP-3 dimerization, providing a glimpse into the first step of coat polymerization. We propose coat polymerization only occurs after cargo engagement, thereby linking cargo sorting with assembly of higher-order coat structures. Additionally, we provide evidence for two amphipathic helices in AP-3, suggesting that AP-3 contributes to membrane deformation during coat assembly. In total, these data provide evidence for the first stages of AP-3-mediated vesicle coat assembly. | |||||||||
| 履歴 |
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構造の表示
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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_45209.map.gz | 217.6 MB | EMDBマップデータ形式 | |
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| ヘッダ (付随情報) | emd-45209-v30.xml emd-45209.xml | 43.9 KB 43.9 KB | 表示 表示 | EMDBヘッダ |
| FSC (解像度算出) | emd_45209_fsc.xml | 13.3 KB | 表示 | FSCデータファイル |
| 画像 | emd_45209.png | 109 KB | ||
| マスクデータ | emd_45209_msk_1.map emd_45209_msk_2.map | 244.1 MB 244.1 MB | マスクマップ | |
| Filedesc metadata | emd-45209.cif.gz | 8.5 KB | ||
| その他 | emd_45209_additional_1.map.gz emd_45209_additional_2.map.gz emd_45209_additional_3.map.gz emd_45209_additional_4.map.gz emd_45209_additional_5.map.gz emd_45209_additional_6.map.gz emd_45209_half_map_1.map.gz emd_45209_half_map_2.map.gz | 121.7 MB 230 MB 1.2 MB 1.2 MB 215.7 MB 3.1 MB 226.6 MB 226.6 MB | ||
| アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-45209 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-45209 | HTTPS FTP |
-関連構造データ
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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 |
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マップ
| ファイル | ダウンロード / ファイル: emd_45209.map.gz / 形式: CCP4 / 大きさ: 244.1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| 注釈 | deepEMHancer sharpened map for Arf1 dimer | ||||||||||||||||||||||||||||||||||||
| 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: 1.058 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
+マスク #1
+マスク #2
+追加マップ: Unsharpened map for Arf1 dimer
+追加マップ: Sharpened map for Arf1 dimer
+追加マップ: Mask for Arf1 dimer
+追加マップ: Refined mask for Arf1 dimer
+追加マップ: Sharpened map for Arf1 dimer
+追加マップ: Map for filtering by local resoluion for Arf1 dimer
+ハーフマップ: Unfiltered half map for Arf1 dimer
+ハーフマップ: Unfiltered half map for Arf1 dimer
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試料の構成要素
-全体 : AP-3 Arf1 dimeric interface, focused refinement
| 全体 | 名称: AP-3 Arf1 dimeric interface, focused refinement |
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| 要素 |
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-超分子 #1: AP-3 Arf1 dimeric interface, focused refinement
| 超分子 | 名称: AP-3 Arf1 dimeric interface, focused refinement / タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#4 |
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| 分子量 | 理論値: 256 KDa |
-分子 #1: AP-3 complex subunit beta-1
| 分子 | 名称: AP-3 complex subunit beta-1 / タイプ: protein_or_peptide / ID: 1 / 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: 77.286523 KDa |
| 組換発現 | 生物種: ![]() |
| 配列 | 文字列: MSSNSFPYNE QSGGGEATEL GQEATSTISP SGAFGLFSSD LKKNEDLKQM LESNKDSAKL DAMKRIVGMI AKGKNASELF PAVVKNVAS KNIEIKKLVY VYLVRYAEEQ QDLALLSIST FQRALKDPNQ LIRASALRVL SSIRVPIIVP IMMLAIKEAS A DLSPYVRK ...文字列: MSSNSFPYNE QSGGGEATEL GQEATSTISP SGAFGLFSSD LKKNEDLKQM LESNKDSAKL DAMKRIVGMI AKGKNASELF PAVVKNVAS KNIEIKKLVY VYLVRYAEEQ QDLALLSIST FQRALKDPNQ LIRASALRVL SSIRVPIIVP IMMLAIKEAS A DLSPYVRK NAAHAIQKLY SLDPEQKEML IEVIEKLLKD KSTLVAGSVV MAFEEVCPDR IDLIHKNYRK LCNLLVDVEE WG QVVIIHM LTRYARTQFV SPWKEGDELE DNGKNFYESD DDQKEKTDKK KKPYTMDPDH RLLIRNTKPL LQSRNAAVVM AVA QLYWHI SPKSEAGIIS KSLVRLLRSN REVQYIVLQN IATMSIQRKG MFEPYLKSFY VRSTDPTMIK TLKLEILTNL ANEA NISTL LREFQTYVKS QDKQFAAATI QTIGRCATNI LEVTDTCLNG LVCLLSNRDE IVVAESVVVI KKLLQMQPAQ HGEII KHMA KLLDSITVPV ARASILWLIG ENCERVPKIA PDVLRKMAKS FTSEDDLVKL QILNLGAKLY LTNSKQTKLL TQYILN LGK YDQNYDIRDR TRFIRQLIVP NVKSGALSKY AKKIFLAQKP APLLESPFKD RDHFQLGTLS HTLNIKATGY LELSNWP EV APDPSVRNVE VIELAKEWTP AGKAKQENSA KKFYSGLEVL FQ UniProtKB: AP-3 complex subunit beta-1 |
-分子 #2: AP-3 complex subunit mu-1
| 分子 | 名称: AP-3 complex subunit mu-1 / 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: 46.989965 KDa |
| 組換発現 | 生物種: ![]() |
| 配列 | 文字列: MIHSLFLINC SGDIFLEKHW KSVVSQSVCD YFFEAQEKAA DVENVPPVIS TPHHYLISIY RDKLFFVSVI QTEVPPLFVI EFLHRVADT FQDYFGECSE AAIKDNVVIV YELLEEMLDN GFPLATESNI LKELIKPPTI LRSVVNSITG SSNVGDTLPT G QLSNIPWR ...文字列: MIHSLFLINC SGDIFLEKHW KSVVSQSVCD YFFEAQEKAA DVENVPPVIS TPHHYLISIY RDKLFFVSVI QTEVPPLFVI EFLHRVADT FQDYFGECSE AAIKDNVVIV YELLEEMLDN GFPLATESNI LKELIKPPTI LRSVVNSITG SSNVGDTLPT G QLSNIPWR RAGVKYTNNE AYFDVVEEID AIIDKSGSTV FAEIQGVIDA CIKLSGMPDL SLSFMNPRLL DDVSFHPCIR FK RWESERV LSFIPPDGNF RLISYRVSSQ NLVAIPVYVK HSISFKENSS CGRFDITIGP KQNMGKTIEG ITVTVHMPKV VLN MNLTPT QGSYTFDPVT KVLTWDVGKI TPQKLPSLKG LVNLQSGAPK PEENPSLNIQ FKIQQLAISG LKVNRLDMYG EKYK PFKGV KYVTKAGKFQ VRT UniProtKB: AP-3 complex subunit mu-1 |
-分子 #3: ADP-ribosylation factor 1
| 分子 | 名称: ADP-ribosylation factor 1 / タイプ: protein_or_peptide / ID: 3 / コピー数: 2 / 光学異性体: LEVO / EC番号: small monomeric GTPase |
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| 由来(天然) | 生物種: Homo sapiens (ヒト) |
| 分子量 | 理論値: 20.775812 KDa |
| 組換発現 | 生物種: ![]() |
| 配列 | 文字列: GNIFANLFKG LFGKKEMRIL MVGLDAAGKT TILYKLKLGE IVTTIPTIGF NVETVEYKNI SFTVWDVGGL DKIRPLWRHY FQNTQGLIF VVDSNDRERV NEAREELMRM LAEDELRDAV LLVFANKQDL PNAMNAAEIT DKLGLHSLRH RNWYIQATCA T SGDGLYEG LDWLSNQLRN QKSL UniProtKB: ADP-ribosylation factor 1 |
-分子 #4: Lysosome-associated membrane glycoprotein 1
| 分子 | 名称: Lysosome-associated membrane glycoprotein 1 / 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: 1.377553 KDa |
| 配列 | 文字列: GRKRSHAGYQ TI UniProtKB: Lysosome-associated membrane glycoprotein 1 |
-分子 #5: MAGNESIUM ION
| 分子 | 名称: MAGNESIUM ION / タイプ: ligand / ID: 5 / コピー数: 2 / 式: MG |
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| 分子量 | 理論値: 24.305 Da |
-分子 #6: GUANOSINE-5'-TRIPHOSPHATE
| 分子 | 名称: GUANOSINE-5'-TRIPHOSPHATE / タイプ: ligand / ID: 6 / コピー数: 2 / 式: GTP |
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| 分子量 | 理論値: 523.18 Da |
| Chemical component information | ![]() ChemComp-GTP: |
-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 | particle |
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試料調製
| 濃度 | 1 mg/mL | |||||||||||||||||||||||||||
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| 緩衝液 | pH: 7.4 構成要素:
詳細: 1x PBS (pH 7.4), 150mM NaCl, 1mM TCEP, 2mM GTP, 5mM EDTA, 10mM MgCl2 | |||||||||||||||||||||||||||
| グリッド | モデル: Quantifoil R1.2/1.3 / 材質: COPPER / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: PLASMA CLEANING / 前処理 - 時間: 200 sec. / 前処理 - 雰囲気: AIR 詳細: Quantifoil Active grids (SPT Labtech) with backside gold coated. Plasma was 12mA for cleaning | |||||||||||||||||||||||||||
| 凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 294 K / 装置: SPOTITON / 詳細: Commercial form of the chameleon (SPT Labtech). | |||||||||||||||||||||||||||
| 詳細 | AP-3 dimer bound to myristoylated Arf1 (Q71L) and LAMP1 on a lipid nanodisc; focused refinement on Arf1 dimeric interface |
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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 (6k x 4k) / 平均電子線量: 53.37 e/Å2 詳細: 2 datasets collected independently and merged for processing |
| 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): 1.4000000000000001 µm 最小 デフォーカス(公称値): 0.4 µm / 倍率(公称値): 81000 |
| 試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
| 実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
-原子モデル構築 1
| 初期モデル | Chain - Source name: AlphaFold / Chain - Initial model type: in silico model |
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| 精密化 | プロトコル: AB INITIO MODEL |
| 得られたモデル | ![]() PDB-9c5a: |
ムービー
コントローラー
万見について




キーワード
Homo sapiens (ヒト)
データ登録者
米国, 1件
引用






































Z (Sec.)
Y (Row.)
X (Col.)






































































































FIELD EMISSION GUN

