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首页> 外文期刊>Biological chemistry >Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition.
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Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition.

机译：由于环境成分中的大分子拥挤效应，SARS-CoV 3CL肽酶活性增加。

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The 3C-like peptidase of the severe acute respiratory syndrome virus (SARS-CoV) is strictly required for viral replication, thus being a potential target for the development of antiviral agents. In contrast to monomeric picornavirus 3C peptidases, SARS-CoV 3CLpro exists in equilibrium between the monomer and dimer forms in solution, and only the dimer is proteolytically active in dilute buffer solutions. In this study, the increase of SARS-CoV 3CLpro peptidase activity in presence of kosmotropic salts and crowding agents is described. The activation followed the Hofmeister series of anions, with two orders of magnitude enhancement in the presence of NaSO, whereas the crowding agents polyethylene glycol and bovine serum albumin increased the hydrolytic rate up to 3 times. Kinetic determinations of the monomer dimer dissociation constant (K(d)) indicated that activation was a result of a more active dimer, without significant changes in K(d) values. The activation was found to be independent of substrate length and was derived from both k(cat) increase and K(m) decrease. The viral peptidase activation described here could be related to the crowded intracellular environment and indicates a further fine-tuning mechanism for biological control, particularly in the microenvironment of the vesicles that are induced in host cells during positive strand RNA virus infection.
机译：病毒复制严格要求使用重症急性呼吸综合征病毒（SARS-CoV）的3C样肽酶，因此是开发抗病毒药物的潜在目标。与单体小核糖核酸病毒3C肽酶相反，SARS-CoV 3CLpro在溶液中的单体和二聚体形式之间处于平衡状态，并且只有二聚体在稀缓冲液中具有蛋白水解活性。在这项研究中，描述了在存在同调性盐和拥挤剂的情况下SARS-CoV 3CLpro肽酶活性的增加。活化过程遵循的是Hofmeister系列阴离子，在NaSO存在下提高了两个数量级，而拥挤剂聚乙二醇和牛血清白蛋白的水解速率提高了3倍。单体二聚体解离常数（K（d））的动力学测定表明，活化是活性更高的二聚体的结果，K（d）值没有明显变化。发现活化与底物长度无关，并且源自k（cat）增加和K（m）减少。本文所述的病毒肽酶激活可能与拥挤的细胞内环境有关，并指示了进一步的微调机制，可用于生物学控制，尤其是在正链RNA病毒感染过程中在宿主细胞中诱导的囊泡微环境中。

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