+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7bc5 | ||||||||||||
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タイトル | Cryo-EM structure of ACP domain from Pichia pastoris fatty acid synthase (FAS) | ||||||||||||
要素 | Fatty acid synthase subunit alpha | ||||||||||||
キーワード | BIOSYNTHETIC PROTEIN / Multienzyme / Complex / Fatty acid / Synthase | ||||||||||||
機能・相同性 | 機能・相同性情報 mitotic nuclear membrane biogenesis / palmitic acid biosynthetic process / fatty-acyl-CoA synthase system / fatty-acyl-CoA synthase activity / fatty acid synthase complex / holo-[acyl-carrier-protein] synthase activity / beta-ketoacyl-[acyl-carrier-protein] synthase I / 3-oxoacyl-[acyl-carrier-protein] reductase / 3-oxoacyl-[acyl-carrier-protein] reductase (NADPH) activity / fatty acid synthase activity ...mitotic nuclear membrane biogenesis / palmitic acid biosynthetic process / fatty-acyl-CoA synthase system / fatty-acyl-CoA synthase activity / fatty acid synthase complex / holo-[acyl-carrier-protein] synthase activity / beta-ketoacyl-[acyl-carrier-protein] synthase I / 3-oxoacyl-[acyl-carrier-protein] reductase / 3-oxoacyl-[acyl-carrier-protein] reductase (NADPH) activity / fatty acid synthase activity / 3-oxoacyl-[acyl-carrier-protein] synthase activity / magnesium ion binding 類似検索 - 分子機能 | ||||||||||||
生物種 | Komagataella phaffii (菌類) | ||||||||||||
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 | Snowden, J.S. / Alzahrani, J. / Sherry, L. / Stacey, M. / Rowlands, D.J. / Ranson, N.A. / Stonehouse, N.J. | ||||||||||||
資金援助 | 英国, サウジアラビア, 3件
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引用 | ジャーナル: Sci Rep / 年: 2021 タイトル: Structural insight into Pichia pastoris fatty acid synthase. 著者: Joseph S Snowden / Jehad Alzahrani / Lee Sherry / Martin Stacey / David J Rowlands / Neil A Ranson / Nicola J Stonehouse / 要旨: Type I fatty acid synthases (FASs) are critical metabolic enzymes which are common targets for bioengineering in the production of biofuels and other products. Serendipitously, we identified FAS as a ...Type I fatty acid synthases (FASs) are critical metabolic enzymes which are common targets for bioengineering in the production of biofuels and other products. Serendipitously, we identified FAS as a contaminant in a cryoEM dataset of virus-like particles (VLPs) purified from P. pastoris, an important model organism and common expression system used in protein production. From these data, we determined the structure of P. pastoris FAS to 3.1 Å resolution. While the overall organisation of the complex was typical of type I FASs, we identified several differences in both structural and enzymatic domains through comparison with the prototypical yeast FAS from S. cerevisiae. Using focussed classification, we were also able to resolve and model the mobile acyl-carrier protein (ACP) domain, which is key for function. Ultimately, the structure reported here will be a useful resource for further efforts to engineer yeast FAS for synthesis of alternate products. | ||||||||||||
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 | 7bc5.cif.gz | 74.4 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7bc5.ent.gz | 34.5 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7bc5.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
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/bc/7bc5 ftp://data.pdbj.org/pub/pdb/validation_reports/bc/7bc5 | HTTPS FTP |
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-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
#1: タンパク質 | 分子量: 206375.734 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Komagataella phaffii (strain GS115 / ATCC 20864) (菌類) 株: GS115 / ATCC 20864 参照: UniProt: C4QY10, fatty-acyl-CoA synthase system, 3-oxoacyl-[acyl-carrier-protein] reductase, beta-ketoacyl-[acyl-carrier-protein] synthase I |
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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: Fatty acid synthase / タイプ: COMPLEX / Entity ID: all / 由来: NATURAL |
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分子量 | 値: 2.6 MDa / 実験値: NO |
由来(天然) | 生物種: Komagataella phaffii GS115 (菌類) |
緩衝液 | pH: 7.4 / 詳細: PBS |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
急速凍結 | 装置: LEICA EM GP / 凍結剤: ETHANE / 湿度: 80 % / 凍結前の試料温度: 281 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 / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 800 nm |
撮影 | 電子線照射量: 60.1 e/Å2 / 検出モード: INTEGRATING フィルム・検出器のモデル: FEI FALCON III (4k x 4k) |
-解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
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: 117012 詳細: Asymmetric reconstruction without alignment was performed on each of two subsets (randomly assigned) from an ACP domain density-containing class after focussed 3D classification. This led to ...詳細: Asymmetric reconstruction without alignment was performed on each of two subsets (randomly assigned) from an ACP domain density-containing class after focussed 3D classification. This led to the generation of two half-maps, which were used to generate a sharpened map from all the data. Note that the number of particles given above includes particles contributing to multiple symmetrically redundant orientations, as particles were generated through focussed 3D classification. 対称性のタイプ: POINT |