Proteomic analysis of Eimeria acervulina sporozoite proteins interaction with duodenal epithelial cells by shotgun LC–MS/MS

Zhang, Zhenchao; Wang, Shuai; Huang, Jingsheng; Liu, Lianrui; Lu, Mingmin; Li, Menghui; Sui, Yuxia; Xu, Lixin; Ren, Yan; Song, Xiaoling; Li, Xiangrui

doi:10.1016/j.molbiopara.2015.09.006

Cited by 12 publications

(13 citation statements)

References 15 publications

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“…Chicken jejunal epithelial cells were isolated using a method described previously [ 11 , 12 ]. Coccidian-free chickens were immerged for 5 min in 70% ethanol after they were slaughtered and the blood was drained.…”

Section: Methodsmentioning

confidence: 99%

Proteomic analysis of protein interactions between Eimeria maxima sporozoites and chicken jejunal epithelial cells by shotgun LC-MS/MS

Huang

Liu

et al. 2018

Parasites Vectors

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BackgroundEimeria maxima initiates infection by invading the jejunal epithelial cells of chicken. However, the proteins involved in invasion remain unknown. The research of the molecules that participate in the interactions between E. maxima sporozoites and host target cells will fill a gap in our understanding of the invasion system of this parasitic pathogen.MethodsIn the present study, chicken jejunal epithelial cells were isolated and cultured in vitro. Western blot was employed to analyze the soluble proteins of E. maxima sporozoites that bound to chicken jejunal epithelial cells. Co-immunoprecipitation (co-IP) assay was used to separate the E. maxima proteins that bound to chicken jejunal epithelial cells. Shotgun LC-MS/MS technique was used for proteomics identification and Gene Ontology was employed for the bioinformatics analysis.ResultsThe results of Western blot analysis showed that four proteins bands from jejunal epithelial cells co-cultured with soluble proteins of E. maxima sporozoites were recognized by the positive sera, with molecular weights of 70, 90, 95 and 130 kDa. The co-IP dilutions were analyzed by shotgun LC-MS/MS. A total of 204 proteins were identified in the E. maxima protein database using the MASCOT search engine. Thirty-five proteins including microneme protein 3 and 7 had more than two unique peptide counts and were annotated using Gene Ontology for molecular function, biological process and cellular localization. The results revealed that of the 35 annotated peptides, 22 (62.86%) were associated with binding activity and 15 (42.86%) were involved in catalytic activity.ConclusionsOur findings provide an insight into the interaction between E. maxima and the corresponding host cells and it is important for the understanding of molecular mechanisms underlying E. maxima invasion.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-2818-4) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Proteomic analysis of protein interactions between Eimeria maxima sporozoites and chicken jejunal epithelial cells by shotgun LC-MS/MS

Huang

Liu

et al. 2018

Parasites Vectors

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“…The existence of a SNF1/ AMPK orthologue was observed in Cryptosporidium parvum (Artz et al 2011) that lacks Krebs cycle and oxidative phosphorylation. In Eimeria acervulina, a putative protein belonging to the SNF1 family was identified and correlated with invasion and evasion mechanisms in chicken duodenal epithelial cells (Zhang et al 2015). Finally, still in the apicomplexa phylum, a potential AMPK homolog (ToxPK1) was observed in Toxoplasma gondii genome with 58 % identity to human AMPK alpha.…”

Section: Snf1/ampk Pathways In Parasitesmentioning

confidence: 99%

AMPK in Pathogens

Mesquita

Moreira

Sampaio‐Marques

et al. 2016

Experientia Supplementum

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During host-pathogen interactions, a complex web of events is crucial for the outcome of infection. Pathogen recognition triggers powerful cellular signaling events that is translated into the induction and maintenance of innate and adaptive host immunity against infection. In opposition, pathogens employ active mechanisms to manipulate host cell regulatory pathways toward their proliferation and survival. Among these, subversion of host cell energy metabolism by pathogens is currently recognized to play an important role in microbial growth and persistence. Extensive studies have documented the role of AMP-activated protein kinase (AMPK) signaling, a central cellular hub involved in the regulation of energy homeostasis, in host-pathogen interactions. Here, we highlight the most recent advances detailing how pathogens hijack cellular metabolism by suppressing or increasing the activity of the host energy sensor AMPK. We also address the role of lower eukaryote AMPK orthologues in the adaptive process to the host microenvironment and their contribution for pathogen survival, differentiation, and growth. Finally, we review the effects of pharmacological or genetic AMPK modulation on pathogen growth and persistence.

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“…Sporozoites from E. acervulina oocysts were purified on DE-52 anion-exchange columns using a previously described protocol [34].…”

Section: Experimental Chickens and Parasitesmentioning

confidence: 99%

“…The duodenal epithelial cells of two-week-old chicks were isolated as previously described [34]. Coccidian-free chicks were emerged in 70% ethanol after they were killed by exsanguination.…”

Section: Isolation and Identification Of Chicken Duodenal Epithelial mentioning

confidence: 99%

Molecular characterization of a potential receptor of Eimeria acervulina microneme protein 3 from chicken duodenal epithelial cells

et al. 2020

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Eimeria acervulina is one of seven Eimeria spp. that can infect chicken duodenal epithelial cells. Eimeria microneme protein 3 (MIC3) plays a vital role in the invasion of host epithelial tissue by the parasite. In this study, we found that chicken (Gallus gallus) ubiquitin conjugating enzyme E2F (UBE2F) could bind to the MIC3 protein of E. acervulina (EaMIC3), as screened using the yeast two-hybrid system, and that it might be the putative receptor protein of EaMIC3. The UBE2F gene was cloned from chicken duodenal epithelial cells. The recombinant protein of UBE2F (rUBE2F) was expressed in E. coli and the reactogenicity of rUBE2F was analyzed by Western blot. Gene sequencing revealed that the opening reading frame (ORF) of UBE2F was 558 base pairs and encoded a protein of 186 amino acids with a molecular weight of 20.46 kDa. The predicted UBE2F protein did not contain signal peptides or a transmembrane region, but had multiple O-glycosylation and phosphorylation sites. A phylogenetic analysis showed that the chicken UBE2F protein is closely related to those of quail and pigeon (Coturnix japonica and Columba livia). A sporozoite invasion-blocking assay showed that antisera against rUBE2F significantly inhibited the invasion of E. acervulina sporozoites in vitro. Animal experiments indicated that the antisera could significantly enhance average body weight gains and reduce mean lesion scores following a challenge with E. acervulina. These results therefore imply that the chicken UBE2F protein might be the target receptor molecule of EaMIC3 that is involved in E. acervulina invasion.

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Proteomic analysis of Eimeria acervulina sporozoite proteins interaction with duodenal epithelial cells by shotgun LC–MS/MS

Cited by 12 publications

References 15 publications

Proteomic analysis of protein interactions between Eimeria maxima sporozoites and chicken jejunal epithelial cells by shotgun LC-MS/MS

Proteomic analysis of protein interactions between Eimeria maxima sporozoites and chicken jejunal epithelial cells by shotgun LC-MS/MS

AMPK in Pathogens

Molecular characterization of a potential receptor of Eimeria acervulina microneme protein 3 from chicken duodenal epithelial cells

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