STRUCTURE OF PSEUDOMONAS-AERUGINOSA ZINC-AZURIN MUTANT ASN47ASP AT 2.4-ANGSTROM RESOLUTION

被引:20
作者
SJOLIN, L
TSAI, LC
LANGER, V
PASCHER, T
KARLSSON, G
NORDLING, M
NAR, H
机构
[1] GOTHENBURG UNIV,DEPT BIOCHEM & BIOPHYS,S-41296 GOTHENBURG,SWEDEN
[2] MAX PLANCK INST BIOCHEM,STRUKTURFORSCH ABT,W-8033 MARTINSRIED,GERMANY
来源
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY | 1993年 / 49卷
关键词
D O I
10.1107/S0907444993005207
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The Pseudomonas aeruginosa azurin mutant Asn47Asp has been isolated, its spectroscopic and kinetic properties characterized, and the X-ray crystal structure of its zinc derivative determined. While the optical and electron paramagnetic resonance spectra as well as the electron-transfer activity of the mutant are very similar to the wild-type values, the Asn47Asp reduction potential is slightly increased by 20 mV. The mutant crystallized in the orthorhombic space group P2(1)2(1)2(1) with cell dimensions a = 57.8, b = 81.5 and c = 112.6 angstrom. There are four molecules in the asymmetric unit, packed as a tetramer which consists of two independent dimers. The zinc site of this mutant structure is similar to the wild-type zinc azurin and, in particular, the metal-binding site is almost identical to the site found in the wild-type zinc-azurin structure [Nar, Huber, Messerschmidt, Filippou, Barth, Jaquinod, Kamp & Canters (1992). Eur. J. Biochem. 205, 1123-11291. The Asp47 side chain at that mutation site takes on a very similar orientation to Asn47 in the wild-type structure preserving the two hydrogen bonds with the neighbouring Thr113 NH and O(gamma)H. Therefore, the increased reduction potential of the mutant is probably a result of an altered charge distribution close to the metal site.
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页码:449 / 457
页数:9
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