Nickel binding to NikA: an additional binding site reconciles spectroscopy, calorimetry and crystallography

Addy C; Ohara M; Kawai FKidera AIkeguchi MFuchigami SOsawa MShimada IPark SYTame JRH

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Nickel binding to NikA: an additional binding site reconciles spectroscopy, calorimetry and crystallography

机译：NikA镍绑定:额外的结合位点和解光谱学、量热法和晶体学

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Intracellular nickel is required by Escherichia coli as a cofactor for a number of enzymes and is necessary for anaerobic respiration. However, high concentrations of nickel are toxic, so both import and export systems have evolved to control the cellular level of the metal. The nik operon in E. coli encodes a nickel-uptake system that includes the periplasmic nickel-binding protein NikA. The crystal structures of wildtype NikA both bound to nickel and in the apo form have been solved previously. The liganded structure appeared to show an unusual interaction between the nickel and the protein in which no direct bonds are formed. The highly unusual nickel coordination suggested by the crystal structure contrasted strongly with earlier X-ray spectroscopic studies. The known nickel-binding site has been probed by extensive mutagenesis and isothermal titration calorimetry and it has been found that even large numbers of disruptive mutations appear to have little effect on the nickel affinity. The crystal structure of a binding-site mutant with nickel bound has been solved and it is found that nickel is bound to two histidine residues at a position distant from the previously characterized binding site. This novel site immediately resolves the conflict between the crystal structures and other biophysical analyses. The physiological relevance of the two binding sites is discussed.

机译：细胞内由大肠镍是必需的杆菌作为辅因子的酶和无氧呼吸的必要条件。高浓度的镍是有毒的,所以两者进出口系统已经进化到控制细胞水平上的金属。大肠杆菌编码nickel-uptake系统包括周质的nickel-binding蛋白质尼卡。绑定到镍和apo形式先前被解决。显示一个不寻常的相互作用没有直接的镍和蛋白质债券形成的。协调建议的晶体结构与早期的x射线形成鲜明对比光谱研究。网站已经被广泛的诱变和探索等温滴定量热法已发现,即使大量的破坏性突变似乎有什么影响镍亲和。结合位点突变与镍绑定解决和发现镍必然两个组氨酸残基位置远离以前的特点结合位点。小说网站立即解决冲突晶体结构和其他之间生物物理分析。讨论了两个结合位点。

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《Acta crystallographica.Section D. Biological crystallography》 |2007年第2期|221-229|共9页
作者
Addy C; Ohara M; Kawai FKidera AIkeguchi MFuchigami SOsawa MShimada IPark SYTame JRH;
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作者单位

Univ Tokyo, Grad Sch Pharmaceut Sci, Dept Chem Phys, Tokyo 1130033, Japan;

Nara Inst Sci & Technol, Grad Sch Mat Sci, Nara 6300192, Japan;

Natl Inst Adv Ind Sci & Technol, Biol Informat Res Ctr, Koto Ku, Tokyo 1350064, JapanYokohama City Univ, Grad Sch Integrated Sci, Yokohama, Kanagawa 2300045, Japan;

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正文语种英语
中图分类化学;
关键词
Escherichia coli; crystal structure; CrystallographyBinding SitesX-ray diffractionUreaseNickel;

机译：结构;CrystallographyBinding SitesX-raydiffractionUreaseNickel;

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机译：Influential factor contributing to the isoform-specific inhibition by aTp of human mitochondrial NaD(p)(+)-dependent malic enzyme - Functional roles of the nucleotide binding site Lys346

Nickel binding to NikA: an additional binding site reconciles spectroscopy, calorimetry and crystallography

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