寄生虫与医学昆虫学报2025，Vol.32Issue(3)：129-137,173,10.DOI:10.3969/j.issn.1005-0507.2025.03.001
微小隐孢子虫亲环蛋白CpCyP1的亚细胞定位及其酶动力学研究
SUBCELLULAR LOCALIZATION AND ENZYMATIC PARAMETERS OF CYCLOPHILIN PROTEIN-1 FROM CRYPTOSPORIDIUM PARVUM
摘要
Abstract
Objective The aim of this study was to characterize the basic molecular and biochemical parameters for a cyclophilin protein in Cryptosporidium parvum called CpCyP1.Methods CpCyP1 expression patterns during the parasite life cycle were evaluated using qRT-PCR with total RNA isolated from different developmental stages of C.parvum.Native CpCyP1 protein in sporozoites was detected using western blot.The localization of CpCyP1 was performed using the immunofluorescence assay,with an affinity-purified rabbit polyclonal antibody against a synthetic peptide.The peptidyl-prolyl cis-trans isomerase(PPIase)activity of His-tagged recombinant CpCyP1 was evaluated using absorbance colorimetry,and the effect of cyclosporin A(CsA)on the activity of CpCyP1 was determined.Results CpCyP1 was expressed in all parasite developmental stages,whereas CpCyP1 was present mainly in the cytosol of sporozoites,meronts,and gamonts.CpCyP1 displayed Michaelis-Menten kinetics towards N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide for its PPIase activity(Km=456.4 μmol/L;Vmax=1.981 U).CsA inhibited PPIase activity,showing lower micromolar inhibitory activity and binding affinity(Kd=5.122 μmol/L;IC50=1.004 μmol/L).Conclusions These results imply that CpCyP1 in the parasite may be the target for the previously reported anti-cryptosporidial efficacy of CsA and suggest that C.parvum cyclophilins could be evaluated as candidate drug targets.关键词
微小隐孢子虫/亲环蛋白/酶动力学/蛋白定位Key words
Cryptosporidium parvum/Cyclophilin/Enzyme kinetics/Protein localization分类
基础医学引用本文复制引用
靳昔蒙,姜鹏,王东强,翟宗振,尹继刚,朱冠..微小隐孢子虫亲环蛋白CpCyP1的亚细胞定位及其酶动力学研究[J].寄生虫与医学昆虫学报,2025,32(3):129-137,173,10.基金项目
"十四五"国家重点研发计划资助(2022YFD1800200) （2022YFD1800200）
