Matthes Nele, Mesters Jeroen R, Coutard Bruno, Canard Bruno, Snijder Eric J, Moll Ralf, Hilgenfeld Rolf
Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck, 23538 Lubeck, Germany.
FEBS Lett. 2006 Jul 24;580(17):4143-9. doi: 10.1016/j.febslet.2006.06.061. Epub 2006 Jun 30.
The non-structural protein Nsp10 of coronaviruses is a small cleavage product of the viral replicase polyprotein that has been implicated in RNA synthesis. Nsp10 of mouse hepatitis virus (MHV) displays an apparent molecular mass of 13-16kDa in reducing SDS-PAGE and analytical gel filtration, while dynamic light scattering suggests the existence of oligomeric forms. Atomic absorption spectroscopy reveals two metal ions per Nsp10 monomer, with a preference for Zn(2+) over Fe(2+/3+) and Co(2+). These are probably bound by two Zn-finger-like motifs. Moreover, MHV Nsp10 interacts with tRNA, single-stranded RNA, double-stranded DNA and, to a lesser extent, single-stranded DNA as shown by gel-shift experiments. The K(d) for tRNA is 2.1+/-0.2 microM.
冠状病毒的非结构蛋白Nsp10是病毒复制酶多聚蛋白的一个小切割产物,与RNA合成有关。在还原SDS-PAGE和分析凝胶过滤中,小鼠肝炎病毒(MHV)的Nsp10显示出明显的分子量为13-16kDa,而动态光散射表明存在寡聚形式。原子吸收光谱显示每个Nsp10单体有两个金属离子,优先结合Zn(2+)而非Fe(2+/3+)和Co(2+)。这些金属离子可能由两个类似锌指的基序结合。此外,凝胶迁移实验表明,MHV Nsp10与tRNA、单链RNA、双链DNA相互作用,与单链DNA的相互作用程度较小。tRNA的解离常数K(d)为2.1±0.2微摩尔。