ジャーナル: J Biol Chem / 年: 2015 タイトル: DNA induces conformational changes in a recombinant human minichromosome maintenance complex. 著者: Emma L Hesketh / Richard P Parker-Manuel / Yuriy Chaban / Rabab Satti / Dawn Coverley / Elena V Orlova / James P J Chong / 要旨: ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in ...ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in higher eukaryotes such as Drosophila, MCM-associated DNA helicase activity has been observed only in the context of a co-purified Cdc45-MCM-GINS complex. Here, we describe the production of the recombinant human MCM (hMCM) complex in Escherichia coli. This protein displays ATP hydrolysis activity and is capable of unwinding duplex DNA. Using single-particle asymmetric EM reconstruction, we demonstrate that recombinant hMCM forms a hexamer that undergoes a conformational change when bound to DNA. Recombinant hMCM produced without post-translational modifications is functional in vitro and provides an important tool for biochemical reconstitution of the human replicative helicase.
ダウンロード / ファイル: emd_2873.map.gz / 形式: CCP4 / 大きさ: 29.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Reconstruction of hMCM bound DNA
ボクセルのサイズ
X=Y=Z: 2.5 Å
密度
表面レベル
登録者による: 0.04 / ムービー #1: 1.6791088
最小 - 最大
-0.75365782 - 5.00809717
平均 (標準偏差)
0.02831108 (±0.2496849)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
-92
-92
-92
サイズ
200
200
200
Spacing
200
200
200
セル
A=B=C: 500.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
2.5
2.5
2.5
M x/y/z
200
200
200
origin x/y/z
0.000
0.000
0.000
length x/y/z
500.000
500.000
500.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
-92
-92
-92
NC/NR/NS
200
200
200
D min/max/mean
-0.754
5.008
0.028
-
添付データ
-
試料の構成要素
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全体 : Recombinant human mini chromosome maintenance complex bound to a ...
全体
名称: Recombinant human mini chromosome maintenance complex bound to a DNA substrate
要素
試料: Recombinant human mini chromosome maintenance complex bound to a DNA substrate
タンパク質・ペプチド: human minichromosome maintenance (2-7) complex
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超分子 #1000: Recombinant human mini chromosome maintenance complex bound to a ...
超分子
名称: Recombinant human mini chromosome maintenance complex bound to a DNA substrate タイプ: sample / ID: 1000 / 集合状態: heterohexamer / Number unique components: 1
分子量
理論値: 567 KDa
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分子 #1: human minichromosome maintenance (2-7) complex