ジャーナル: EMBO J / 年: 2003 タイトル: Nucleotide-induced conformations in the neck region of dimeric kinesin. 著者: Georgios Skiniotis / Thomas Surrey / Stephan Altmann / Heinz Gross / Young-Hwa Song / Eckhard Mandelkow / Andreas Hoenger / 要旨: The neck region of kinesin constitutes a key component in the enzyme's walking mechanism. Here we applied cryoelectron microscopy and image reconstruction to investigate the location of the kinesin ...The neck region of kinesin constitutes a key component in the enzyme's walking mechanism. Here we applied cryoelectron microscopy and image reconstruction to investigate the location of the kinesin neck in dimeric and monomeric constructs complexed to microtubules. To this end we enhanced the visibility of this region by engineering an SH3 domain into the transition between neck linker and neck coiled coil. The resulting chimeric kinesin constructs remained functional as verified by physiology assays. In the presence of AMP-PNP the SH3 domains allowed us to identify the position of the neck in a well defined conformation and revealed its high flexibility in the absence of nucleotide. We show here the double-headed binding of dimeric kinesin along the same protofilament, which is characterized by the opposite directionality of neck linkers. In this configuration the neck coiled coil appears fully zipped. The position of the neck region in dimeric constructs is not affected by the presence of the tubulin C-termini as confirmed by subtilisin treatment of microtubules prior to motor decoration.
ダウンロード / ファイル: emd_1039.map.gz / 形式: CCP4 / 大きさ: 3.7 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
rat kinesin dimer with an SH3 domain cloned within the neck region -rKS379-, complexed to subtilisin treated microtubules in the presence of AMP-PNP
ボクセルのサイズ
X=Y=Z: 5.526 Å
密度
表面レベル
1: 63.600000000000001 / ムービー #1: 54.4705846
最小 - 最大
0.0 - 100.0
平均 (標準偏差)
40.3611 (±10.633699999999999)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
100
100
100
Spacing
100
100
100
セル
A=B=C: 552.6 Å α=β=γ: 90 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
5.526
5.526
5.526
M x/y/z
100
100
100
origin x/y/z
0.000
0.000
0.000
length x/y/z
552.600
552.600
552.600
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
100
100
100
D min/max/mean
0.000
100.000
40.361
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添付データ
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試料の構成要素
-
全体 : Rat kinesin dimer with an SH3 domain complexed to subtilisin trea...
全体
名称: Rat kinesin dimer with an SH3 domain complexed to subtilisin treated microtubules in the presence of AMP-PNP.
要素
試料: Rat kinesin dimer with an SH3 domain complexed to subtilisin treated microtubules in the presence of AMP-PNP.
タンパク質・ペプチド: rat kinesin
タンパク質・ペプチド: tubulin
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超分子 #1000: Rat kinesin dimer with an SH3 domain complexed to subtilisin trea...
超分子
名称: Rat kinesin dimer with an SH3 domain complexed to subtilisin treated microtubules in the presence of AMP-PNP. タイプ: sample / ID: 1000 / 集合状態: dimer / Number unique components: 2