Characterization of a Species-Specific Insulinase-Like Protease in Cryptosporidium parvum

Autor: Shijing Zhang, Yuping Wang, Haizhen Wu, Na Li, Jianlin Jiang, Yaqiong Guo, Yaoyu Feng, Lihua Xiao
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Frontiers in Microbiology, Vol 10 (2019)
Druh dokumentu: article
ISSN: 1664-302X
DOI: 10.3389/fmicb.2019.00354
Popis: Cryptosporidium parvum is an intracellular protozoan that can cause severe diarrhea in humans and various mammals. Results of a comparative genomic analysis indicated that genes encoding two C. parvum-specific insulinase-like proteases (INS19 and INS20), cgd6_5510 and cgd6_5520, are lost in many Cryptosporidium species. In this study, we provided evidence indicating that cgd6_5510 and cgd6_5520 are fragments of a full gene (cgd6_5520-5510) encoding one insulinase-like protease (INS20-19) that is similar in structure to classic insulinases. We expressed cgd6_5510 in Escherichia coli for antiserum preparation and found the protein (INS19) that was partially degraded. A ~180 kDa protein of INS20-19 was specifically recognized by the polyclonal anti-INS19 antiserum in sporozoite lysate. We observed that INS20-19 is likely a protein with high expression in the apical region of sporozoites, and neutralization of the protein led to a partial reduction of parasite load in HCT-8 and MDBK cell cultures at 24 h. Taken together, our findings support the involvement of INS20-19 in the invasion or early developmental process of C. parvum.
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