Comparative analyses and structural insights of new class glutathione transferases in Cryptosporidium species

Abstract

Cryptosporidiosis, caused by protozoan parasites of the genus Cryptosporidium, is estimated to rank as a leading cause in the global burden of neglected zoonotic parasitic diseases. This diarrheal disease is the second leading cause of death in children under 5 years of age. Based on the C. parvum transcriptome data, glutathione transferase (GST) has been suggested as a drug target against this pathogen. GSTs are diverse multifunctional proteins involved in cellular defense and detoxification in organisms and help pathogens to alleviate chemical and environmental stress. In this study, we performed genome-wide data mining, identification, classification and in silico structural analysis of GSTs in fifteen Cryptosporidium species. The study revealed the presence three GSTs in each of the Cryptosporidium species analyzed in the study. Based on the percentage identity and comprehensive comparative phylogenetic analysis, we assigned Cryptosporidium species GSTs to three new GST classes, named Vega (ϑ), Gamma (γ) and Psi (ψ). The study also revealed an atypical thioredoxin-like fold in the C. parvum GST1 of the Vega class, whereas C. parvum GST2 of the Gamma class and C. melagridis GST3 of the Psi class has a typical thioredoxin-like fold in the N-terminal region. This study reports the first comparative analysis of GSTs in Cryptosporidium species.

Caseinolytic Proteins (Clp) in the Genus Klebsiella: Special Focus on ClpK

“…ClpK has been identified as an ATPase since it consists of structural motifs associated with the hydrolysis of ATP [ 14 , 16 , 17 , 19 ]. To our knowledge, the ability of ClpK to hydrolyse ATP in vitro has not been reported to date.…”

“…Walker A forms the floor of the nucleotide-binding pocket and binds the ATP phosphate. Walker B positions cations to bind metals that play a role in ATP catalysis [ 14 , 16 , 17 , 19 ].…”

A novel, stain-free, natural auto-fluorescent signal, Sig M, identified from cytometric and transcriptomic analysis of infectivity of Cryptosporidium hominis and Cryptosporidium parvum

“…(2018) ( Supplementary Table S4 .). The extra eight Cryptosporidium genes were selected based on the analysis of potential drug targets from recent studies ( Manjunatha et al., 2017 ; Baragaña et al., 2019 ; Castellanos-Gonzalez et al., 2019 ; Su et al., 2019 ; Xu et al., 2019 ; Zhang et al., 2019 ; Mfeka et al., 2020 ). The gene IDs from the study were further cross-referenced with data from the CryptoDB database ( http://cryptodb.org ), from which the corresponding mRNA sequences were extracted.…”

“…The panel used consisted of 144 genes: 40 human genes frequently expressed in human cell lines selected as control genes to ascertain if the assay was functioning as expected and 104 Cryptosporidium genes extracted from the analysis of previous RNA-seq studies, with 48 thought to be expressed intracellularly and 48 extracellularly according to data from Matos et al, 2019 (Supplementary Tables S2, S3) and crossreferenced with Lippuner et al (2018) (Supplementary Table S4.). The extra eight Cryptosporidium genes were selected based on the analysis of potential drug targets from recent studies (Manjunatha et al, 2017;Baragaña et al, 2019;Castellanos-Gonzalez et al, 2019;Su et al, 2019;Xu et al, 2019;Zhang et al, 2019;Mfeka et al, 2020). The gene IDs from the study were further cross-referenced with data from the CryptoDB database (http://cryptodb.org), from which the corresponding mRNA sequences were extracted.…”

“…We assessed the expression of Cryptosporidium and human host genes to attempt to gain a better understanding of the expression of genes at various stages of the life cycle of this pathogen. To that end, a panel of genes from previous transcriptomic studies ( Lippuner et al., 2018 ; Matos et al., 2019 ) and a selection of potential drug targets ( Manjunatha et al., 2017 ; Baragaña et al., 2019 ; Castellanos-Gonzalez et al., 2019 ; Su et al., 2019 ; Xu et al., 2019 ; Zhang et al., 2019 ; Mfeka et al., 2020 ) were chosen, and their expression was assessed.…”