Enzymatic activity of the SARS coronavirus main proteinase dimer.

Enzymatic activity of the SARS coronavirus main proteinase dimer.
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SARS冠状病毒主蛋白酶二聚体的酶促活性。

DOI:
10.1016/j.febslet.2006.04.004
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发表时间:
2006-05-15
期刊:
影响因子:
3.5
通讯作者:
Mangel WF
Mangel WF
中科院分区:
生物学3区
文献类型:
--
作者:
Graziano V;McGrath WJ;DeGruccio AM;Dunn JJ;Mangel WF

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SARS冠状病毒主要蛋白酶二聚体的酶活性通过灵敏的定量测定来表征。新的荧光底物(Ala‐Arg‐Leu‐Gln‐NH)2‐罗丹明含有严重急性呼吸综合征冠状病毒(SARS CoV)主要蛋白酶共有切割序列和罗丹明110(已知最易检测的化合物之一)作为报告基团。在没有纯化标签的情况下克隆酶的基因,在大肠杆菌中表达并纯化酶。SARS冠状病毒主要蛋白酶二聚体的酶活性可以在低pM浓度下容易地检测到。该酶具有较高的Km值,对离子强度和还原剂异常敏感。
The enzymatic activity of the SARS coronavirus main proteinase dimer was characterized by a sensitive, quantitative assay. The new, fluorogenic substrate, (Ala‐Arg‐Leu‐Gln‐NH)2‐Rhodamine, contained a severe acute respiratory syndrome coronavirus (SARS CoV) main proteinase consensus cleavage sequence and Rhodamine 110, one of the most detectable compounds known, as the reporter group. The gene for the enzyme was cloned in the absence of purification tags, expressed in Escherichia coli and the enzyme purified. Enzyme activity from the SARS CoV main proteinase dimer could readily be detected at low pM concentrations. The enzyme exhibited a high K m, and is unusually sensitive to ionic strength and reducing agents.
冠状病毒主蛋白酶的结构揭示了甲over依蛋白酶折叠与额外的α-螺旋结构域的组合。
DOI: 10.1093/emboj/cdf327
发表时间: 2002-07-01
期刊: EMBO JOURNAL
影响因子: 11.4
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