贺双意¹, 孔佳^1,2, 王媚媚¹, 米立志¹, 秦晓红^1,3
AuthorsHTML:贺双意¹, 孔佳^1,2, 王媚媚¹, 米立志¹, 秦晓红^1,3
AuthorsListE:He Shuangyi¹, Kong Jia^1,2, Wang Meimei¹, Mi Lizhi¹, Qin Xiaohong^1,3
AuthorsHTMLE:He Shuangyi¹, Kong Jia^1,2, Wang Meimei¹, Mi Lizhi¹, Qin Xiaohong^1,3
Unit:1. 天津大学生命科学学院，天津 300072；2. 天津大学化工学院，天津 300350；3. 南开大学药物化学生物学国家重点实验室，天津 300071
Unit_EngLish:1. School of Life Science, Tianjin University, Tianjin 300072, China
2. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China
3. State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300071, China
Abstract_Chinese:SAMHD1蛋白是一种天然免疫限制因子, 利用其dNTP水解酶活性或核酸酶活性抑制病毒的复制, 具有广谱抗病毒功能．利用生物信息学分析该蛋白家族序列保守性, 并借助HPLC层析色谱分析与酶活性测定和酶标仪方法, 对人源、猪源和鸡源三物种SAMHD1蛋白的酶活性进行了比较．比对发现SAMHD1蛋白在活性空腔、变构位点及磷酸化位点等处序列上具有高度的保守性; 而且发现猪源和鸡源SAMHD1蛋白同样具有dNTP水解酶和核酸酶活性; 3种蛋白中, 人源SAMHD1蛋白dNTP水解酶活性最高, 猪源蛋白核酸酶活性最低．这一结果为研究SAMHD1蛋白家族的结构功能关系及为畜牧业优良品种的开发提供启示．
Abstract_English:Sterile α-motif/histidine-aspartate domain-containing protein 1(SAMHD1)，an innate immunological factor，can restrict viral replication by using its deoxynucleotide triphosphohydrolase(dNTPase)or nuclease activities，thus having a broad spectrum of antiviral functions. The sequence conservativeness of human，porcine，and chicken SAMHD1s was analyzed by bioinformatics，and their enzymatic activities were compared by using high performance liquid chromatography(HPLC) and a fluorescent-based enzymatic assay. It is found that the allosteric，nucleotide binding，and phosphorylation sites of SAMHD1 were highly conservative in sequence. Although all the SAMHD1s from the three different species had dNTPase and nuclease activities，they did show difference in the measured activities. Among them，human SAMHD1 had the highest dNTPase activity，and the porcine SAMHD1 had the lowest nuclease activity. This study provides information for understanding SAMHD1 structure-activity relationships and the development of new breeds in the livestock industry.
Keyword_Chinese:SAMHD1; 物种; 蛋白表达与纯化; 三磷酸水解酶活性; 核酸酶活性
Keywords_English:SAMHD1; species; protein expression and purification; dNTPase activity; nuclease activity

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