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データを開く
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基本情報
登録情報 | データベース: SASBDB / ID: SASDD46 |
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![]() | Artificially designed de novo protein esPN-Block (HL4) heterocomplex, e1s2 (HL4)
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生物種 | artificially designed de novo protein (人工物) |
![]() | ![]() タイトル: Self-Assembling Supramolecular Nanostructures Constructed from de Novo Extender Protein Nanobuilding Blocks. 著者: Naoya Kobayashi / Kouichi Inano / Kenji Sasahara / Takaaki Sato / Keisuke Miyazawa / Takeshi Fukuma / Michael H Hecht / Chihong Song / Kazuyoshi Murata / Ryoichi Arai / ![]() ![]() 要旨: The design of novel proteins that self-assemble into supramolecular complexes is important for development in nanobiotechnology and synthetic biology. Recently, we designed and created a protein ...The design of novel proteins that self-assemble into supramolecular complexes is important for development in nanobiotechnology and synthetic biology. Recently, we designed and created a protein nanobuilding block (PN-Block), WA20-foldon, by fusing an intermolecularly folded dimeric de novo WA20 protein and a trimeric foldon domain of T4 phage fibritin (Kobayashi et al., J. Am. Chem. Soc. 2015, 137, 11285). WA20-foldon formed several types of self-assembling nanoarchitectures in multiples of 6-mers, including a barrel-like hexamer and a tetrahedron-like dodecamer. In this study, to construct chain-like polymeric nanostructures, we designed de novo extender protein nanobuilding blocks (ePN-Blocks) by tandemly fusing two de novo binary-patterned WA20 proteins with various linkers. The ePN-Blocks with long helical linkers or flexible linkers were expressed in soluble fractions of Escherichia coli, and the purified ePN-Blocks were analyzed by native PAGE, size exclusion chromatography-multiangle light scattering (SEC-MALS), small-angle X-ray scattering (SAXS), and transmission electron microscopy. These results suggest formation of various structural homo-oligomers. Subsequently, we reconstructed hetero-oligomeric complexes from extender and stopper PN-Blocks by denaturation and refolding. The present SEC-MALS and SAXS analyses show that extender and stopper PN-Block (esPN-Block) heterocomplexes formed different types of extended chain-like conformations depending on their linker types. Moreover, atomic force microscopy imaging in liquid suggests that the esPN-Block heterocomplexes with metal ions further self-assembled into supramolecular nanostructures on mica surfaces. Taken together, the present data demonstrate that the design and construction of self-assembling PN-Blocks using de novo proteins is a useful strategy for building polymeric nanoarchitectures of supramolecular protein complexes. |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
-モデル
モデル #1899 | ![]() タイプ: dummy / ダミー原子の半径: 2.75 A / 対称性: P1 / カイ2乗値: 0.004 / P-value: 0.678914 ![]() |
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モデル #1919 | ![]() タイプ: atomic / カイ2乗値: 0.036 ![]() |
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試料
![]() | 名称: Artificially designed de novo protein esPN-Block (HL4) heterocomplex, e1s2 (HL4) 試料濃度: 2.6 mg/ml / Entity id: 1028 / 1029 |
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バッファ | 名称: 20 mM HEPES, 100 mM NaCl, 200 mM ArgHCl, 10% glycerol pH: 7.5 |
要素 #1028 | 名称: ePN-Block (HL4) / タイプ: protein / 記述: extender PN-Block (HL4) / 分子量: 27.484 / 分子数: 1 / 由来: artificially designed de novo protein 配列: MYGKLNKLVE HIKELLQQLN KNWHRHQGNL HDMNQQMEQL FQEFQHFMQG NQDDGKLQNM IHEMQQFMNQ VDNHLQSESD TVHHFHNKLQ ELMNNFHHLV HRKLAEAAAK EAAAKEAAAK EAAAKAAAMY GKLNKLVEHI KELLQQLNKN WHRHQGNLHD MNQQMEQLFQ ...配列: MYGKLNKLVE HIKELLQQLN KNWHRHQGNL HDMNQQMEQL FQEFQHFMQG NQDDGKLQNM IHEMQQFMNQ VDNHLQSESD TVHHFHNKLQ ELMNNFHHLV HRKLAEAAAK EAAAKEAAAK EAAAKAAAMY GKLNKLVEHI KELLQQLNKN WHRHQGNLHD MNQQMEQLFQ EFQHFMQGNQ DDGKLQNMIH EMQQFMNQVD NHLQSESDTV HHFHNKLQEL MNNFHHLVHR |
要素 #1029 | 名称: sPN-Block (WA20) / タイプ: protein / 記述: stopper PN-Block (WA20) / 分子量: 12.547 / 分子数: 2 / 由来: artificially designed de novo protein 配列: MYGKLNKLVE HIKELLQQLN KNWHRHQGNL HDMNQQMEQL FQEFQHFMQG NQDDGKLQNM IHEMQQFMNQ VDNHLQSESD TVHHFHNKLQ ELMNNFHHLV HR |
-実験情報
ビーム | 設備名称: Photon Factory (PF), High Energy Acceleration Research Organization (KEK) BL-10C 地域: Tsukuba / 国: Japan ![]() | ||||||||||||||||||||||||||||||
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検出器 | 名称: Pilatus3 2M / Pixsize x: 0.172 mm | ||||||||||||||||||||||||||||||
スキャン | 測定日: 2014年12月19日 / セル温度: 20 °C / 照射時間: 5 sec. / フレーム数: 30 / 単位: 1/nm /
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