ジャーナル: Science / 年: 2009 タイトル: Proteome organization in a genome-reduced bacterium. 著者: Sebastian Kühner / Vera van Noort / Matthew J Betts / Alejandra Leo-Macias / Claire Batisse / Michaela Rode / Takuji Yamada / Tobias Maier / Samuel Bader / Pedro Beltran-Alvarez / Daniel ...著者: Sebastian Kühner / Vera van Noort / Matthew J Betts / Alejandra Leo-Macias / Claire Batisse / Michaela Rode / Takuji Yamada / Tobias Maier / Samuel Bader / Pedro Beltran-Alvarez / Daniel Castaño-Diez / Wei-Hua Chen / Damien Devos / Marc Güell / Tomas Norambuena / Ines Racke / Vladimir Rybin / Alexander Schmidt / Eva Yus / Ruedi Aebersold / Richard Herrmann / Bettina Böttcher / Achilleas S Frangakis / Robert B Russell / Luis Serrano / Peer Bork / Anne-Claude Gavin / 要旨: The genome of Mycoplasma pneumoniae is among the smallest found in self-replicating organisms. To study the basic principles of bacterial proteome organization, we used tandem affinity purification- ...The genome of Mycoplasma pneumoniae is among the smallest found in self-replicating organisms. To study the basic principles of bacterial proteome organization, we used tandem affinity purification-mass spectrometry (TAP-MS) in a proteome-wide screen. The analysis revealed 62 homomultimeric and 116 heteromultimeric soluble protein complexes, of which the majority are novel. About a third of the heteromultimeric complexes show higher levels of proteome organization, including assembly into larger, multiprotein complex entities, suggesting sequential steps in biological processes, and extensive sharing of components, implying protein multifunctionality. Incorporation of structural models for 484 proteins, single-particle electron microscopy, and cellular electron tomograms provided supporting structural details for this proteome organization. The data set provides a blueprint of the minimal cellular machinery required for life.
ダウンロード / ファイル: emd_1639.map.gz / 形式: CCP4 / 大きさ: 1001 KB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
RNA-polymerase from Mycoplasma pneumoniae
ボクセルのサイズ
X=Y=Z: 5.2 Å
密度
表面レベル
登録者による: 5.0 / ムービー #1: 5
最小 - 最大
0.0 - 48.7149
平均 (標準偏差)
0.205414 (±1.90841)
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
64
64
64
Spacing
64
64
64
セル
A=B=C: 332.8 Å α=β=γ: 90 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
5.2
5.2
5.2
M x/y/z
64
64
64
origin x/y/z
0.000
0.000
0.000
length x/y/z
332.800
332.800
332.800
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-64
-64
-64
NX/NY/NZ
128
128
128
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
64
64
64
D min/max/mean
0.000
48.715
0.205
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添付データ
-
試料の構成要素
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全体 : RNA-polymerase of Mycoplasma pneumoniae
全体
名称: RNA-polymerase of Mycoplasma pneumoniae
要素
試料: RNA-polymerase of Mycoplasma pneumoniae
タンパク質・ペプチド: RNA-polymerase
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超分子 #1000: RNA-polymerase of Mycoplasma pneumoniae
超分子
名称: RNA-polymerase of Mycoplasma pneumoniae / タイプ: sample / ID: 1000 / 詳細: Sample was fixed following the GRAFIX protocol / Number unique components: 1
タイプ: NEGATIVE 詳細: Grids were prepared by sandwich negative stain the sample was adsorbed to carbon on mica and floated on 1 % uranyl acetate the carbon was picked up with an uncoated grid then a second piece ...詳細: Grids were prepared by sandwich negative stain the sample was adsorbed to carbon on mica and floated on 1 % uranyl acetate the carbon was picked up with an uncoated grid then a second piece of carbon, which was also floated on 1% uranyl acetate, was picked up with the same grid sandwiching the sample between the two layers of carbon
グリッド
詳細: 400 mesh copper grid
凍結
凍結剤: NONE / 装置: OTHER
-
電子顕微鏡法
顕微鏡
FEI/PHILIPS CM200FEG
アライメント法
Legacy - 非点収差: Corrected at 200000 times magnification on graininess of carbon
撮影
カテゴリ: CCD / フィルム・検出器のモデル: GENERIC CCD / デジタル化 - サンプリング間隔: 14.22 µm / 実像数: 96 詳細: Images were recorded on CCD, no scanning sampling step size was adjusted to calibrated image size ビット/ピクセル: 12
Tilt angle min
0
Tilt angle max
0
電子線
加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2 mm / 倍率(公称値): 27500
試料ステージ
試料ホルダー: Eucentric / 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC
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画像解析
最終 再構成
想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 25.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: IMAGIC, SPIDER, EMAN 詳細: Spider option BP 32F Back Projection - 3D, Sampled, Interpolated in Fourier space 使用した粒子像数: 5324