文献类型：期刊文献

英文题名：Comparative Characterization of CpCDPK1 and CpCDPK9, Two Potential Drug Targets against Cryptosporidiosis

作者：Su, Jiayuan[1];Shen, Yiting[1];Li, Na[2,3];Li, Yu[4];Zhang, Ziding[4];Xiao, Lihua[2,3];Guo, Yaqiong[1,2];Feng, Yaoyu[1,2,3]

机构：[1]East China Univ Sci & Technol, Sch Resource & Environm Engn, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]South China Agr Univ, Coll Vet Med, Ctr Emerging & Zoonot Dis, Guangzhou 510642, Peoples R China;[3]Guangdong Lab Lingnan Modern Agr, Guangzhou 510642, Peoples R China;[4]China Agr Univ, Coll Biol Sci, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China

年份：2022

卷号：10

期号：2

外文期刊名：MICROORGANISMS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000824121100005)】；

基金：This research was funded by the National Natural Science Foundation of China (grant numbers 32030109 and U1901208); the 111 Project (grant number D20008); and the Innovation Team Project of Guangdong University (grant number 2019KCXTD001).

语种：英文

外文关键词：Cryptosporidium parvum; calcium-dependent protein kinase; protein expression; invasion; inhibitor

摘要：As the invasion, egress, and growth of Cryptosporidium spp. are regulated by the calcium ion, calcium-dependent protein kinases (CDPKs) are considered potential drug targets against these pathogens. In this study, we expressed CpCDPK1 of Cryptosporidium parvum encoded by the cgd3_920 gene and CpCDPK9 encoded by the the cgd7_1260 gene in Escherichia coli, and we conducted some comparative studies with quantitative PCR, immunofluorescence staining, and in vitro neutralization assays. By immunofluorescence microscopy, CpCDPK1 was expressed over the entirety of the sporozoites, while CpCDPK9 was mainly expressed in the apical region. The expression of the cgd3_920 gene was the highest at 12 h of the in vitro culture, whereas the expression of the cgd7_1260 gene peaked between 2 h and 6 h. Polyclonal antibodies against these two CpCDPK proteins had similar neutralization efficiency on C. parvum growth, reaching approximately 40%. Of the 50 candidate compounds from the molecular docking of CpCDPK1, 10 had significant in vitro anti-cryptosporidial effects, but only one inhibited enzyme activity. For CpCDPK9, five of the forty-five candidate compounds showed significant in vitro anti-cryptosporidial effects. Results obtained from this study suggest that CpCDPK1 and CpCDPK9 might function differently in C. parvum infection.