ジャーナル: J Mol Biol / 年: 2006 タイトル: Human kinetochore-associated kinesin CENP-E visualized at 17 A resolution bound to microtubules. 著者: E Neumann / I Garcia-Saez / S DeBonis / R H Wade / F Kozielski / J F Conway / 要旨: The highly dynamic process of cell division is effected, in part, by molecular motors that generate the forces necessary for its enactment. Several members of the kinesin superfamily of motor ...The highly dynamic process of cell division is effected, in part, by molecular motors that generate the forces necessary for its enactment. Several members of the kinesin superfamily of motor proteins are implicated in mitosis, such as CENP-E, which plays essential roles in cell division, including association with the kinetochore to stabilize attachment of chromosomes to microtubules prior to and during their separation. Neither the functional assembly state of CENP-E nor its direction of motion along the polar microtubule are certain. To determine the mode of interaction between CENP-E and microtubules, we have used cryo-electron microscopy to visualize CENP-E motor domains complexed with microtubules and calculated a density map of the complex to 17 A resolution by combining helical and single-particle reconstruction methods. The interface between the motor domain and microtubules was modeled by docking atomic-resolution models of the subunits into the cryoEM density map. Our results support a plus end motion for CENP-E, consistent with features of the crystallographic structure. Despite considerable functional differences from the monomeric transporter kinesin KIF1A and the oppositely directed ncd kinesin, CENP-E appears to share many features of the intermolecular interactions, suggesting that differences in motor function are governed by small variations in the loops at the microtubule interface.
ダウンロード / ファイル: emd_1309.map.gz / 形式: CCP4 / 大きさ: 2 MB / タイプ: IMAGE STORED AS SIGNED BYTE
注釈
Kinetochore-associated kinesin CENP-E motors bound to microtubules
ボクセルのサイズ
X=Y=Z: 3.56 Å
密度
表面レベル
1: 87.0 / ムービー #1: -35
最小 - 最大
-128.0 - 127.0
平均 (標準偏差)
-87.432599999999994 (±41.033900000000003)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
Z
X
Y
Origin
-63
-63
-63
サイズ
128
128
128
Spacing
128
128
128
セル
A=B=C: 455.68 Å α=β=γ: 90 °
CCP4マップ ヘッダ情報:
mode
envelope stored as signed bytes (from -128 lowest to 127 highest)
Å/pix. X/Y/Z
3.56
3.56
3.56
M x/y/z
128
128
128
origin x/y/z
0.000
0.000
0.000
length x/y/z
455.680
455.680
455.680
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-63
-63
-63
NX/NY/NZ
128
128
128
MAP C/R/S
3
1
2
start NC/NR/NS
-63
-63
-63
NC/NR/NS
128
128
128
D min/max/mean
-128.000
127.000
-87.433
-
添付データ
-
試料の構成要素
-
全体 : Human Kinetochore-associated Kinesin CENP-E bound to Microtubules
全体
名称: Human Kinetochore-associated Kinesin CENP-E bound to Microtubules
要素
試料: Human Kinetochore-associated Kinesin CENP-E bound to Microtubules
タンパク質・ペプチド: Polymerized Tubulin
タンパク質・ペプチド: Human kinetochore-associated kinesin CENP-E motors
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超分子 #1000: Human Kinetochore-associated Kinesin CENP-E bound to Microtubules
超分子
名称: Human Kinetochore-associated Kinesin CENP-E bound to Microtubules タイプ: sample / ID: 1000 集合状態: One CENP-E motor domain binds to a tubulin heterodimer Number unique components: 2
アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 17.0 Å / 解像度の算出法: FSC 0.33 CUT-OFF / ソフトウェア - 名称: MRC and Spider
CTF補正
詳細: CTFMIX
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原子モデル構築 1
ソフトウェア
名称: Situs, URO
詳細
The cryoEM map and the atomic structures of CENP-E and the tubulin dimer were visualized together using the program "O". An initial manual fit was straightforward, and subsequently refined using CoLoRes from the SITUS package, and URO.