- EMDB-1797: Proteomic characterization of archaeal ribosomes reveals the pres... -
+
データを開く
IDまたはキーワード:
読み込み中...
-
基本情報
登録情報
データベース: EMDB / ID: EMD-1797
タイトル
Proteomic characterization of archaeal ribosomes reveals the presence of novel archaeal-specific ribosomal proteins. 50S ribosomal subunit of Sulfolobus acidocaldarius
マップデータ
This is a 50S ribosomal subunit of Sulfolobus acidocaldarius
ジャーナル: J Mol Biol / 年: 2011 タイトル: Proteomic characterization of archaeal ribosomes reveals the presence of novel archaeal-specific ribosomal proteins. 著者: Viter Márquez / Thomas Fröhlich / Jean-Paul Armache / Daniel Sohmen / Alexandra Dönhöfer / Aleksandra Mikolajka / Otto Berninghausen / Michael Thomm / Roland Beckmann / Georg J Arnold / Daniel N Wilson / 要旨: Protein synthesis occurs in macromolecular particles called ribosomes. All ribosomes are composed of RNA and proteins. While the protein composition of bacterial and eukaryotic ribosomes has been ...Protein synthesis occurs in macromolecular particles called ribosomes. All ribosomes are composed of RNA and proteins. While the protein composition of bacterial and eukaryotic ribosomes has been well-characterized, a systematic analysis of archaeal ribosomes has been lacking. Here we report the first comprehensive two-dimensional PAGE and mass spectrometry analysis of archaeal ribosomes isolated from the thermophilic Pyrobaculum aerophilum and the thermoacidophilic Sulfolobus acidocaldarius Crenarchaeota. Our analysis identified all 66 ribosomal proteins (r-proteins) of the P. aerophilum small and large subunits, as well as all but two (62 of 64; 97%) r-proteins of the S. acidocaldarius small and large subunits that are predicted genomically. Some r-proteins were identified with one or two lysine methylations and N-terminal acetylations. In addition, we identify three hypothetical proteins that appear to be bona fide r-proteins of the S. acidocaldarius large subunit. Dissociation of r-proteins from the S. acidocaldarius large subunit indicates that the novel r-proteins establish tighter interactions with the large subunit than some integral r-proteins. Furthermore, cryo electron microscopy reconstructions of the S. acidocaldarius and P. aerophilum 50S subunits allow for a tentative localization of the binding site of the novel r-proteins. This study illustrates not only the potential diversity of the archaeal ribosomes but also the necessity to experimentally analyze the archaeal ribosomes to ascertain their protein composition. The discovery of novel archaeal r-proteins and factors may be the first step to understanding how archaeal ribosomes cope with extreme environmental conditions.
pH: 7.5 詳細: 20 mM Hepes pH 7.5, 10 mM Mg(OAc)2, 30 mM NH4OAc, 4 mM beta-Mercaptoethanol
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: OTHER / 詳細: Vitrification instrument: Vitrobot 手法: Blot for 10 seconds before plunging, use 2 layers of filter paper
-
電子顕微鏡法
顕微鏡
FEI TECNAI 12
撮影
カテゴリ: CCD / フィルム・検出器のモデル: FEI EAGLE (2k x 2k) / デジタル化 - サンプリング間隔: 3.308 µm / 実像数: 99 / 平均電子線量: 20 e/Å2 / 詳細: Data collected on CCD