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基本情報
登録情報 | データベース: PDB / ID: 5ai7 | ||||||
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タイトル | ParM doublet model | ||||||
![]() | PLASMID SEGREGATION PROTEIN PARM | ||||||
![]() | MOTOR PROTEIN / PARM / ACTIN-LIKE PROTEIN / DOUBLET / ANTIPARALLEL | ||||||
機能・相同性 | Plasmid segregation protein ParM/StbA / : / Plasmid segregation protein ParM, N-terminal / Plasmid segregation protein ParM, C-terminal / ParM-like / plasmid partitioning / ATPase, nucleotide binding domain / identical protein binding / Plasmid segregation protein ParM![]() | ||||||
生物種 | ![]() ![]() | ||||||
手法 | 電子顕微鏡法 / らせん対称体再構成法 / クライオ電子顕微鏡法 | ||||||
Model type details | CA ATOMS ONLY, CHAIN A, B, C, D, E, F, G, H, I, J, K, L, M, N | ||||||
![]() | Bharat, T.A.M. / Murshudov, G.N. / Sachse, C. / Lowe, J. | ||||||
![]() | ![]() タイトル: Structures of actin-like ParM filaments show architecture of plasmid-segregating spindles. 著者: Tanmay A M Bharat / Garib N Murshudov / Carsten Sachse / Jan Löwe / ![]() ![]() 要旨: Active segregation of Escherichia coli low-copy-number plasmid R1 involves formation of a bipolar spindle made of left-handed double-helical actin-like ParM filaments. ParR links the filaments with ...Active segregation of Escherichia coli low-copy-number plasmid R1 involves formation of a bipolar spindle made of left-handed double-helical actin-like ParM filaments. ParR links the filaments with centromeric parC plasmid DNA, while facilitating the addition of subunits to ParM filaments. Growing ParMRC spindles push sister plasmids to the cell poles. Here, using modern electron cryomicroscopy methods, we investigate the structures and arrangements of ParM filaments in vitro and in cells, revealing at near-atomic resolution how subunits and filaments come together to produce the simplest known mitotic machinery. To understand the mechanism of dynamic instability, we determine structures of ParM filaments in different nucleotide states. The structure of filaments bound to the ATP analogue AMPPNP is determined at 4.3 Å resolution and refined. The ParM filament structure shows strong longitudinal interfaces and weaker lateral interactions. Also using electron cryomicroscopy, we reconstruct ParM doublets forming antiparallel spindles. Finally, with whole-cell electron cryotomography, we show that doublets are abundant in bacterial cells containing low-copy-number plasmids with the ParMRC locus, leading to an asynchronous model of R1 plasmid segregation. | ||||||
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構造ビューア | 分子: ![]() ![]() |
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PDBx/mmCIF形式 | ![]() | 131.3 KB | 表示 | ![]() |
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-検証レポート
文書・要旨 | ![]() | 622.9 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 625.4 KB | 表示 | |
XML形式データ | ![]() | 35.8 KB | 表示 | |
CIF形式データ | ![]() | 68.7 KB | 表示 | |
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-関連構造データ
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リンク
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集合体
登録構造単位 | ![]()
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要素
#1: タンパク質 | 分子量: 35633.223 Da / 分子数: 14 / 由来タイプ: 組換発現 詳細: THIS IS A MODEL OF THE PARM DOUBLET BASED ON CRYO-EM DATA. ONLY CARBON-ALPHAS HAVE BEEN DEPOSITED. 由来: (組換発現) ![]() ![]() ![]() ![]() |
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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: FILAMENT / 3次元再構成法: らせん対称体再構成法 |
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試料調製
構成要素 | 名称: PARM ANTIPARALLEL DOUBLET / タイプ: COMPLEX |
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緩衝液 | 名称: 50 MM TRIS-HCL, 100 MM KCL, AND 1 MM MGCL2, 2% PEG 6000 pH: 7 詳細: 50 MM TRIS-HCL, 100 MM KCL, AND 1 MM MGCL2, 2% PEG 6000 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | 詳細: CARBON |
急速凍結 | 詳細: VIROBOT MARK IV (FEI) |
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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: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 6000 nm / 最小 デフォーカス(公称値): 3000 nm / Cs: 2.7 mm |
撮影 | 電子線照射量: 0.3 e/Å2 フィルム・検出器のモデル: FEI FALCON II (4k x 4k) |
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解析
EMソフトウェア | 名称: SPRING / カテゴリ: 3次元再構成 / 詳細: SPRING (DESFOSSES AND SACHSE) | ||||||||||||
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3次元再構成 | 手法: CRYO-EM ALIGNMENT 詳細: THIS IS A MODEL OF THE PARM ANTIPARALLEL ANTIPARALLEL DOUBLET BASED ON CRYOEM DATA. TWO COPIES OF THE PARM AMPPNP CRYO-EM FILAMENT STRUCTURE WERE PLACED IN AN ANTIPARALLEL ORIENTATION. ...詳細: THIS IS A MODEL OF THE PARM ANTIPARALLEL ANTIPARALLEL DOUBLET BASED ON CRYOEM DATA. TWO COPIES OF THE PARM AMPPNP CRYO-EM FILAMENT STRUCTURE WERE PLACED IN AN ANTIPARALLEL ORIENTATION. ANTIPARALLEL ASSIGNMENT WAS DONE USING CRYO-EM ALIGNMENT. THIS IS A MODEL OF THE PARM ANTIPARALLEL DOUBLET CONSTRUCTED USING CRYO-EM DATA. ONLY C-ALPHAS HAVE BEEN DEPOSITED. 対称性のタイプ: HELICAL | ||||||||||||
精密化ステップ | サイクル: LAST
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