| Project/Area Number |
60470153
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
結晶学
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| Research Institution | Institute for Protein Research, Osaka University |
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Principal Investigator |
KATSUBE Yukiteru Inst. for Protein Research, Osaka Univ. Prof, 蛋白質研究所, 教授 (20032013)
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| Co-Investigator(Kenkyū-buntansha) |
HATA Yasuo Inst,For Protein Research, Osaka Univ. Assist., 蛋白質研究所, 助手 (10127277)
TANAKA Nobuo Inst. for Protein Research, OsakaUniv. Assist. Prof, 蛋白質研究所, 助教授 (50032024)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1985: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Superoxide Dismutase / Protein Crystal Structure / バレル構造 / スーパーオキシドジスムターゼ / タンパク質のX線結晶解析 |
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| Research Abstract |
The complete amino acid sequence of the Cu,Zn-superoxide dismutase(Cu-Zn-SOD) from spinach has been determined on the basis of peptides by brCN cleavage and by proteolyses with some proteases. This is a first sequence determination of the Cu,Zn-SOD. The enzyme is dimer consisting of two identical subunits. Each subunit consists of a total of 154 amino acid residues with an un-blocked N-terminal alanine residue. The crystallization was carried out by gradual addition of ammonium sulfate powder to the protein solution ajusted to pH7.8 with potassium phosphate buffer. Many heavy-atom reagents were tested to get the heavy-atom derivatives available for the X- ray work. Finally,two derivatives obtained by K_3 UO_2 F_5 and K_2 Pt (NO_2)_4 were available for the X-ray analysis. An wlwctron density map at 2 A resolution was computed by using the multiple isomorphous replacement method supplemented by anomalous scattering effect due to the uranyl ions. This map is good enough to a main chain tracing. The subunit structure of the spinach SOD is composed of the barrel with eight <beta>-strands and thrdd loops. The longest loop, in Whish 35 residues are contained, is connected to a <beta>-strand by a S-S bond.
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