Pepsin-like aspartic protease (Sc-ASP155) cloning, molecular characterization and gene expression analysis in developmental stages of nematode Steinernema carpocapsae

Balasubramanian, N; Nascimento, Gisela; Ferreira, Ricardo Melo; Martínez, Mònica; Simões, Nélson

doi:10.1016/j.gene.2012.03.062

Cited by 17 publications

(23 citation statements)

References 41 publications

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“…Proteinases released by T. spiralis play an essential role in parasite invasion [11,12]. Several serine proteinases have been identified and demonstrated to be involved in various adaptive functions, such as tissue invasion and immune evasion, [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of a Trichinella spiralis aspartic protease and its facilitation role in larval invasion of host intestinal epithelial cells

Liu

Bai

et al. 2020

PLoS Negl Trop Dis

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Background T. spiralis aspartic protease has been identified in excretion/secretion (ES) proteins, but its roles in larval invasion are unclear. The aim of this study was to characterize T. spiralis aspartic protease-2 (TsASP2) and assess its roles in T. spiralis invasion into intestinal epithelial cells (IECs) using RNAi. Methodology/Principal findings Recombinant TsASP2 (rTsASP2) was expressed and purified. The native TsASP2 of 43 kDa was recognized by anti-rTsASP2 serum in all worm stages except newborn larvae (NBL), and qPCR indicated that TsASP2 transcription was highest at the stage of intestinal infective larvae (IIL). IFA results confirmed that TsASP2 was located in the hindgut, midgut and muscle cells of muscle larvae (ML) and IIL and intrauterine embryos of the female adult worm (AW), but not in NBL. rTsASP2 cleaved several host proteins (human hemoglobin (Hb), mouse Hb, collagen and IgM). The proteolytic activity of rTsASP2 was host-specific, as it hydrolyzed mouse Hb more efficiently than human Hb. The enzymatic activity of rTsASP2 was significantly inhibited by pepstatin A. The expression levels of TsASP2 mRNA and protein were significantly suppressed by RNAi with 5 μM TsASP2-specific siRNA. Native aspartic protease activity in ML crude proteins was reduced to 54.82% after transfection with siRNA. Larval invasion of IECs was promoted by rTsASP2 and inhibited by anti-rTsASP2 serum and siRNA. Furthermore, cell monolayer damage due to larval invasion was obviously alleviated when siRNA-treated larvae were used. The adult worm burden, length of adult worms and female fecundity were clearly reduced in mice challenged using siRNA-treated ML relative to the PBS group, Conclusions rTsASP2 possesses the enzymatic activity of native aspartic protease and facilitates T. spiralis invasion of host IECs.

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Section: Introductionmentioning

confidence: 99%

Molecular characterization of a Trichinella spiralis aspartic protease and its facilitation role in larval invasion of host intestinal epithelial cells

Liu

Bai

et al. 2020

PLoS Negl Trop Dis

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“…In most cases, they associate with symbiotic bacteria, which they release once inside their insect host [1]. Both nematode-derived [2] and bacterial factors [3] interfere to varying extent with host immune reactions often resulting in the ultimate death of the insect [1]. During recent years, EPNs have increasingly been used to probe the insect immune system and identify genes and factors that have the potential to protect against EPN infections [4].…”

Section: Introductionmentioning

confidence: 99%

Genome-Wide Transcriptional Analysis of <b><i>Drosophila</i></b> Larvae Infected by Entomopathogenic Nematodes Shows Involvement of Complement, Recognition and Extracellular Matrix Proteins

et al. 2013

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Heterorhabditis bacteriophora is an entomopathogenic nematode (EPN) which infects its host by accessing the hemolymph where it releases endosymbiotic bacteria of the species Photorhabdus luminescens. We performed a genome-wide transcriptional analysis of the Drosophila response to EPN infection at the time point at which the nematodes reached the hemolymph either via the cuticle or the gut and the bacteria had started to multiply. Many of the most strongly induced genes have been implicated in immune responses in other infection models. Mapping of the complete set of differentially regulated genes showed the hallmarks of a wound response, but also identified a large fraction of EPN-specific transcripts. Several genes identified by transcriptome profiling or their homologues play protective roles during nematode infections. Genes that positively contribute to controlling nematobacterial infections encode: a homolog of thioester-containing complement protein 3, a basement membrane component (glutactin), a recognition protein (GNBP-like 3) and possibly several small peptides. Of note is that several of these genes have not previously been implicated in immune responses.

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“… 36 , 37 Aspartic proteases could be used to digest the host haemoglobin in parasitic nematodes, such as Brugia malayi , 38 T. spiralis , 39 , 40 and Steinernema carpocapsae . 41 In our study, putative aspartic proteases were found to be secreted from A. cantonensis young adults and detected by 2-DE and LC-MS/MS analysis.…”

Section: Discussionmentioning

confidence: 60%

Proteomic analysis of excretory-secretory products from young adults of Angiostrongylus cantonensis

Chen

Cheng

et al. 2019

Mem. Inst. Oswaldo Cruz

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BACKGROUND Angiostrongyliasis is caused by the nematode Angiostrongylus cantonensis and can lead to eosinophilic meningitis and meningoencephalitis in humans. The young adult worms play central pathogenic roles in the central nervous system (CNS); however, the underlying mechanism is unclear. Excretory-secretory products (ESPs) are good investigation targets for studying the relationship between a host and its parasite. OBJECTIVES We aimed to profile, identify, and characterise the proteins in the ESPs of A. cantonensis young adults. METHODS The ESPs of young adult worms were collected from culture medium after incubation ranging from 24 to 96 h. Proteomic and bioinformatics analyses were performed to characterise the ESPs. FINDINGS A total of 51 spots were identified, and the highly expressed proteins included two protein disulphide isomerases, one calreticulin, and three uncharacterised proteins. Subsequently, approximately 254 proteins were identified in the ESPs of A. cantonensis young adults via liquid chromatography-mass spectrometry (LC-MS/MS) analysis, and these were further classified according to their characteristics and biological functions. Finally, we identified the immunoreactive proteins from a reference map of ESPs from A. cantonensis young adults. Approximately eight proteins were identified, including a protein disulphide isomerase, a putative aspartic protease, annexin, and five uncharacterised proteins. The study established and identified protein reference maps for the ESPs of A. cantonensis young adults. MAIN CONCLUSIONS The identified proteins may be potential targets for the development of diagnostic or therapeutic agents for human angiostrongyliasis.

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Pepsin-like aspartic protease (Sc-ASP155) cloning, molecular characterization and gene expression analysis in developmental stages of nematode Steinernema carpocapsae

Cited by 17 publications

References 41 publications

Molecular characterization of a Trichinella spiralis aspartic protease and its facilitation role in larval invasion of host intestinal epithelial cells

Molecular characterization of a Trichinella spiralis aspartic protease and its facilitation role in larval invasion of host intestinal epithelial cells

Genome-Wide Transcriptional Analysis of <b><i>Drosophila</i></b> Larvae Infected by Entomopathogenic Nematodes Shows Involvement of Complement, Recognition and Extracellular Matrix Proteins

Proteomic analysis of excretory-secretory products from young adults of Angiostrongylus cantonensis

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