Cloning and analysis of a Trichinella pseudospiralis muscle larva secreted serine protease gene

Cwiklinski, Krystyna; Meskill, Diana; Robinson, Mark W.; Pozio, E; Appleton, Judith A.; Connolly, Bernadette

doi:10.1016/j.vetpar.2008.10.036

Cited by 15 publications

(9 citation statements)

References 18 publications

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“…These results indicated that TspSP-1.2 contributes to the larval invasion of host intestinal epithelial cells and could be a potential vaccine candidate against T. spiralis infection [ 9 ]. A similar protein (TppSP-1) from Trichinella pseudospiralis muscle larvae was identified by Cwiklinski et al [ 10 ]. Analysis of the deduced amino acid sequence found that the histidine residue of the catalytic triad in TsSP-1 was replaced with an arginine residue in the TppSP-1.…”

Section: Parasitic Nematode Serine Proteasesmentioning

confidence: 89%

“…Analysis of the deduced amino acid sequence found that the histidine residue of the catalytic triad in TsSP-1 was replaced with an arginine residue in the TppSP-1. This could lead to the loss of proteolytic activity, and the role in the T. pseudospiralis -host interaction needs further research [ 10 ]. Trap et al [ 11 ] identified another putative serine protease by screening a library from T. spiralis adult-newborn larvae mixed stage with a radioisotope-labelled DNA probe.…”

Section: Parasitic Nematode Serine Proteasesmentioning

confidence: 99%

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Serine Proteases of Parasitic Helminths

et al. 2015

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Serine proteases form one of the most important families of enzymes and perform significant functions in a broad range of biological processes, such as intra- and extracellular protein metabolism, digestion, blood coagulation, regulation of development, and fertilization. A number of serine proteases have been identified in parasitic helminths that have putative roles in parasite development and nutrition, host tissues and cell invasion, anticoagulation, and immune evasion. In this review, we described the serine proteases that have been identified in parasitic helminths, including nematodes (Trichinella spiralis, T. pseudospiralis, Trichuris muris, Anisakis simplex, Ascaris suum, Onchocerca volvulus, O. lienalis, Brugia malayi, Ancylostoma caninum, and Steinernema carpocapsae), cestodes (Spirometra mansoni, Echinococcus granulosus, and Schistocephalus solidus), and trematodes (Fasciola hepatica, F. gigantica, and Schistosoma mansoni). Moreover, the possible biological functions of these serine proteases in the endogenous biological phenomena of these parasites and in the host-parasite interaction were also discussed.

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Section: Parasitic Nematode Serine Proteasesmentioning

confidence: 89%

Section: Parasitic Nematode Serine Proteasesmentioning

confidence: 99%

Serine Proteases of Parasitic Helminths

et al. 2015

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“…Several secreted serine proteases have been identified among T . spiralis ES proteins, including the 69 kDa putative serine protease TsSerP (two trypsin-like domains), the 45 kDa serine protease TspSP-1, and a 35.5 kDa serine protease [ 9 , 11 , 16 – 18 ]. Most of these have strong antigenicity, specific antibodies for them are easily detected experimentally in infected animal sera, and they have one or two trypsin like domains [ 9 , 11 , 16 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of a host collagen inducing factor from the excretory secretory proteins of Trichinella spiralis

Park¹,

Kim²,

Cho³

et al. 2018

PLoS Negl Trop Dis

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BackgroundIn a previous study, we found that Trichinella spiralis muscle larva excretory and secretory proteins (ES-P) most likely activate collagen synthesis via TGF-β/Smad signaling, and this event could influence collagen capsule formation.Methodology/Principal findingsIn order to identify the specific collagen inducing factor, ES-P was fractionated by a Superdex 200 10/300 GL column. We obtained three large fractions, F1, F2, and F3, but only F3 had collagen gene inducing ability. After immunoscreening, 10 collagen inducing factor candidates were identified. Among them, TS 15–1 and TS 15–2 were identical to the putative trypsin of T. spiralis. The deduced TS 15–1 (M.W. = 72 kDa) had two conserved catalytic motifs, an N-terminal Tryp_SPc domain (TS 15-1n) and a C-terminal Tryp_SPc domain (TS 15-1c). To determine their collagen inducing ability, recombinant proteins (rTS 15-1n and rTS 15-1c) were produced using the pET-28a expression system. TS 15–1 is highly expressed during the muscle larval stage and has strong antigenicity. We determined that rTS 15-1c could elevate collagen I via activation of the TGF-β1 signaling pathway in vitro and in vivo.Conclusion/SignificanceIn conclusion, we identified a host collagen inducing factor from T. spiralis ES-P using immunoscreening and demonstrated its molecular characteristics and functions.

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“…In S. carpocapsae, preliminary assays also pointed out the presence of serine proteases in ESPs. This finding led us to search for genes encoding serine proteases during the S. car- Independently of intron number in vertebrate and invertebrate serine protease genes, the ending of a particular intron is commonly conserved six nucleotides before the Ser 195 codon (44). This feature is also conserved in the sc-sp-1 gene.…”

Section: Discussionmentioning

confidence: 99%

Serine Protease-mediated Host Invasion by the Parasitic Nematode Steinernema carpocapsae

Toubarro

Lucena-Robles

Nascimento

et al. 2010

Journal of Biological Chemistry

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Steinernema carpocapsae is an insect parasitic nematode used in biological control, which infects insects penetrating by mouth and anus and invading the hemocoelium through the midgut wall. Invasion has been described as a key factor in nematode virulence and suggested to be mediated by proteases. A serine protease cDNA from the parasitic stage was sequenced (sc-sp-1); the recombinant protein was produced in an Escherichia coli system, and a native protein was purified from the secreted products. Both proteins were confirmed by mass spectrometry to be encoded by the sc-sp-1 gene. Sc-SP-1 has a pI of 8.7, a molecular mass of 27.3 kDa, a catalytic efficiency of 22.2 ؋ 10 4 s ؊1 M ؊1 against N-succinyl-Ala-Ala-Pro-Phe-pNA, and is inhibited by chymostatin (IC 0.07) and PMSF (IC 0.73). Sc-SP-1 belongs to the chymotrypsin family, based on sequence and biochemical analysis. Only the nematode parasitic stage expressed sc-sp-1. These nematodes in the midgut lumen, prepared to invade the insect hemocoelium, expressed higher levels than those already in the hemocoelium. Moreover, parasitic nematode sense insect peritrophic membrane and hemolymph more quickly than they do other tissues, which initiates sc-sp-1 expression. Ex vivo, Sc-SP-1 was able to bind to insect midgut epithelium and to cause cell detachment from basal lamina. In vitro, Sc-SP-1 formed holes in an artificial membrane model (Matrigel), whereas Sc-SP-1 treated with PMSF did not, very likely because it hydrolyzes matrix glycoproteins. These findings highlight the S. carpocapsae-invasive process that is a key step in the parasitism thus opening new perspectives for improving nematode virulence to use in biological control.

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Cloning and analysis of a Trichinella pseudospiralis muscle larva secreted serine protease gene

Cited by 15 publications

References 18 publications

Serine Proteases of Parasitic Helminths

Serine Proteases of Parasitic Helminths

Identification of a host collagen inducing factor from the excretory secretory proteins of Trichinella spiralis

Serine Protease-mediated Host Invasion by the Parasitic Nematode Steinernema carpocapsae

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