SUBCELLULAR LOCALIZATION AND ENZYMATIC PARAMETERS OF CYCLOPHILIN PROTEIN-1 FROM CRYPTOSPORIDIUM PARVUM
10.3969/j.issn.1005-0507.2025.03.001
- VernacularTitle: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
- Author:
Xi-Meng JIN
1
;
Peng JIANG
;
Dong-Qiang WANG
;
Zong-Zhen ZHAI
;
Ji-Gang YIN
;
Guan ZHU
Author Information
1. 人畜共患传染病重症诊治全国重点实验室,吉林大学人兽共患病研究所/动物医学学院,长春 130062
- Keywords:
Cryptosporidium parvum;
Cyclophilin;
Enzyme kinetics;
Protein localization
- From:
Acta Parasitologica et Medica Entomologica Sinica
2025;32(3):129-137,173
- CountryChina
- Language:Chinese
-
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.
