Architecture and Principle of Formation of the Baseplate of Bacteriophage T4
| Project/Area Number |
13680736
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
ARISAKA Fumio Grad. Sch. Biosci. & Bioeng. Assoc. Professor, 大学院・生命理工学研究科, 助教授 (80133768)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | tail lysozyme / lysozyme / association equilibrium / β-helix / bacteriophage / X-ray crystallography / infection / 感染 / 分子シャペロン / 繊維状蛋白質 / 熱変性 / グアニジン塩酸変性 / 分子集合 / リゾチーム / 収縮性尾部 / 感染機構 / X線結晶解析 / 超遠心分析 |
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| Research Abstract |
Crystal structure of the tail lysozyme, gp5, which is a structurally essential component of the baseplate and also possesses lysozyme activity has been determined to the resolution of 2.9 A. Gp5 of which the C-terminus had His-Tag was bound to Ni-NTA agarose gel and the column was then used as an affinity chromatograph to isolate separately prepared Seleno-Met substituted gp27 which is another component of the ″hub″of the baseplate for phase determination.
Isolated complex from the Ni-NTA agarose column had the subunit composition of (gp5*)^3(gp5C)^3(gp27)^3, where gp5* is the mature tail lysozyme, gp5C is the C-terminal domain. The overall shape of the complex appears a "torch", where trimer gp27 and the N-terminal domain form the cup, the lysozyme domain forms the rim, and the C-terminal domain assumes the "stem". The three domains of gp5 are connected by two linkers with 44 residues (linker 1) and 49 residues (linker 2). The N-terminal domain possesses a so-called OB motif (OB stands for oligonucleotide/oligosaccharide binding) which is a five-stranded beta-barrel. The lysozyme domain is similar to T4 lysozyme with the rms of 1.1 A. The C-terminal domain forms a long prism with the length of 110 A and the width of 28 A. The N-terminal 46 residues in the C-terminal domain form 5-stranded anti-parallel β-sheets. The three beta sheets, from each subunit, form a short prism. The remaining part forms a triple-stranded β-helix. The cross-section of the β-helix reveals three identical sequence of VXGXXXX from each polypeptide chain with a kink at glycine residue which forms the vertice of each triangle. The β-strands form parallel beta sheets. The β-helix is a very stable structure which is resistant to denaturation by SDS at room temperature.
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Report
(3 results)
Research Products
(22 results)
