Mirzayan C, Wimmer E
Department of Molecular Microbiology, State University of New York at Stony Brook 11794.
Virology. 1994 Feb 15;199(1):176-87. doi: 10.1006/viro.1994.1110.
Poliovirus 2C is a nonstructural polypeptide proposed to function in viral RNA replication. Poliovirus 2C is a member of a rapidly expanding family of proteins containing a consensus for nucleotide binding (NTP-B). Site-directed mutagenesis of conserved residues in the consensus A and B sites have suggested a functional role for the NTP-B motif in viral RNA replication and proliferation of poliovirus. We have expressed wildtype 2C and a 2C mutant, carrying a single amino acid exchange in the NTP-B motif A (Lys135Gln) using the baculovirus system. Both wildtype and mutant proteins are membrane associated. Following membrane solubilization, we have purified wildtype and mutant proteins to near homogeneity using conventional chromatography. We present biochemical evidence that wildtype 2C copurifies with an ATPase activity that is absent in the mutant preparation.
脊髓灰质炎病毒2C是一种非结构多肽，被认为在病毒RNA复制中发挥作用。脊髓灰质炎病毒2C是一个迅速扩展的蛋白质家族的成员，该家族含有核苷酸结合（NTP-B）的共有序列。对共有序列A和B位点保守残基的定点诱变表明，NTP-B基序在脊髓灰质炎病毒的病毒RNA复制和增殖中具有功能作用。我们使用杆状病毒系统表达了野生型2C和一种2C突变体，该突变体在NTP-B基序A中携带单个氨基酸交换（Lys135Gln）。野生型和突变型蛋白均与膜相关。膜溶解后，我们使用常规色谱法将野生型和突变型蛋白纯化至接近均一。我们提供了生化证据，表明野生型2C与一种ATP酶活性共纯化，而该活性在突变体制备中不存在。
