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蝎毒活性肽BmK AngM1具有良好的镇痛活性,但目前对其抗炎活性的研究尚未有报道。本研究利用Escherichia coli BL21 trx B(DE3)对BmK AngM1进行了重组表达,利用IMPACTTM-TWIN系统建立了重组BmK AngM1(rBmK AngM1)的纯化方法,并对r BmK AngM1进行了抗炎活性检测。为了进一步提高其抗炎活性,本研究对BmK AngM1功能结构域中可能的活性位点(Y5、Y42和R58)进行了定点突变。结果表明,rBmK AngM1及其突变体均具有显著的抗炎活性,其中单突变体R58N及双突变体Y5F/R58N、Y42F/R58N与野生型蛋白相比,抗炎活性均有明显提高。由此推测,58位残基在BmK AngM1的抗炎活性中起重要作用。本研究为通过蛋白质工程设计改造BmK AngM1以提高其药理活性奠定了基础。
Scorpion venom active peptide BmK AngM1 has a good analgesic activity, but its anti-inflammatory activity has not been reported. In this study, BmK AngM1 was recombinantly expressed in Escherichia coli BL21 trx B (DE3). The recombinant BmK AngM1 (rBmK AngM1) was purified by IMPACTTM-TWIN system and the anti-inflammatory activity of rBmK AngM1 was tested. In order to further enhance its anti-inflammatory activity, we mutated the possible active sites (Y5, Y42 and R58) in the functional domain of BmK AngM1. The results showed that rBmK AngM1 and its mutants all had significant anti-inflammatory activity. The anti-inflammatory activity of rBmK AngM1 and its mutants was significantly higher than that of wild-type R58N and double mutants Y5F / R58N and Y42F / R58N. Therefore, it is speculated that the 58th residue plays an important role in the anti-inflammatory activity of BmK AngM1. This study laid the foundation for improving the pharmacological activity of BmK AngM1 through the engineering of protein engineering.