Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium.
Journal
PLoS neglected tropical diseases
ISSN: 1935-2735
Titre abrégé: PLoS Negl Trop Dis
Pays: United States
ID NLM: 101291488
Informations de publication
Date de publication:
09 2023
09 2023
Historique:
received:
09
06
2023
accepted:
30
08
2023
revised:
21
09
2023
medline:
25
9
2023
pubmed:
11
9
2023
entrez:
11
9
2023
Statut:
epublish
Résumé
A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteristics, and investigate its role on larval invasion of gut epithelium during T. spiralis infection. TsSPc has a functional domain of serine proteinase, and its tertiary structure consists of three amino acid residues (His88, Asp139 and Ser229) forming a pocket like functional domain. Recombinant TsSPc (rTsSPc) was expressed and purified. The rTsSPc has good immunogenicity. On Western blot analysis, rTsSPc was recognized by infection serum and anti-rTsSPc serum, natural TsSPc in crude and ES antigens was identified by anti-rTsSPc serum. The results of qPCR, Western blot and indirect immunofluorescence test (IIFT) showed that TsSPc was expressed at diverse stage worms, and mainly localized at cuticle, stichosome and intrauterine embryos of this nematode. The rTsSPc had enzymatic activity of native serine protease, which hydrolyzed the substrate BAEE, casein and collagen I. After site directed mutation of enzymatic active sites of TsSPc, its antigenicity did not change but the enzyme activity was fully lost. rTsSPc specifically bound to intestinal epithelium cells (IECs) and the binding sites were mainly localized in cell membrane and cytoplasm. rTsSPc accelerated larval invasion of IECs, whereas anti-rTsSPc antibodies and TsSPc-specific dsRNA obviously hindered larval invasion. TsSPc was a surface and secretory proteinase of the parasite, participated in larval invasion of gut epithelium, and may be considered as a candidate vaccine target molecule against Trichinella intrusion and infection.
Sections du résumé
BACKGROUND
A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteristics, and investigate its role on larval invasion of gut epithelium during T. spiralis infection.
METHODOLOGY/PRINCIPAL FINDINGS
TsSPc has a functional domain of serine proteinase, and its tertiary structure consists of three amino acid residues (His88, Asp139 and Ser229) forming a pocket like functional domain. Recombinant TsSPc (rTsSPc) was expressed and purified. The rTsSPc has good immunogenicity. On Western blot analysis, rTsSPc was recognized by infection serum and anti-rTsSPc serum, natural TsSPc in crude and ES antigens was identified by anti-rTsSPc serum. The results of qPCR, Western blot and indirect immunofluorescence test (IIFT) showed that TsSPc was expressed at diverse stage worms, and mainly localized at cuticle, stichosome and intrauterine embryos of this nematode. The rTsSPc had enzymatic activity of native serine protease, which hydrolyzed the substrate BAEE, casein and collagen I. After site directed mutation of enzymatic active sites of TsSPc, its antigenicity did not change but the enzyme activity was fully lost. rTsSPc specifically bound to intestinal epithelium cells (IECs) and the binding sites were mainly localized in cell membrane and cytoplasm. rTsSPc accelerated larval invasion of IECs, whereas anti-rTsSPc antibodies and TsSPc-specific dsRNA obviously hindered larval invasion.
CONCLUSIONS
TsSPc was a surface and secretory proteinase of the parasite, participated in larval invasion of gut epithelium, and may be considered as a candidate vaccine target molecule against Trichinella intrusion and infection.
Identifiants
pubmed: 37695792
doi: 10.1371/journal.pntd.0011629
pii: PNTD-D-23-00721
pmc: PMC10513378
doi:
Substances chimiques
Serine Proteases
EC 3.4.-
Serine Endopeptidases
EC 3.4.21.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0011629Informations de copyright
Copyright: © 2023 Song et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
Res Vet Sci. 2020 Jun;130:110-117
pubmed: 32171999
Parasitol Res. 2015 Dec;114(12):4637-44
pubmed: 26342828
BMC Biotechnol. 2014 Dec 28;14:109
pubmed: 25543846
Int J Parasitol. 2008 Feb;38(2):149-55
pubmed: 18048044
Vet Res Forum. 2021;12(3):291-295
pubmed: 34815839
J Proteomics. 2012 Apr 18;75(8):2375-83
pubmed: 22348823
Nat Genet. 2011 Mar;43(3):228-35
pubmed: 21336279
Exp Parasitol. 2015 Dec;159:264-73
pubmed: 26545353
Exp Parasitol. 2015 Jun;153:29-38
pubmed: 25733024
Acta Trop. 2022 May;229:106388
pubmed: 35231417
Vet Res. 2018 Sep 6;49(1):87
pubmed: 30189894
Methods Mol Biol. 2017;1626:13-24
pubmed: 28608196
Vet Parasitol. 2009 Feb 23;159(3-4):272-5
pubmed: 19091476
Food Res Int. 2018 Jul;109:380-386
pubmed: 29803463
Front Microbiol. 2017 May 31;8:986
pubmed: 28620363
Int J Parasitol. 2007 Feb;37(2):139-48
pubmed: 17007860
Acta Trop. 2022 Feb;226:106263
pubmed: 34879232
Acta Trop. 2015 Oct;150:79-86
pubmed: 26184560
Parasit Vectors. 2018 Sep 6;11(1):499
pubmed: 30189888
Food Microbiol. 2017 Jun;64:65-71
pubmed: 28213036
Front Microbiol. 2021 Mar 17;12:634185
pubmed: 33815318
PLoS Negl Trop Dis. 2015 Jun 05;9(6):e0003807
pubmed: 26046773
Acta Trop. 2021 Apr;216:105825
pubmed: 33421420
Vet Parasitol. 2021 Sep;297:109160
pubmed: 32522393
PLoS Negl Trop Dis. 2018 May 14;12(5):e0006485
pubmed: 29758030
Vet Res. 2020 Sep 25;51(1):125
pubmed: 32988413
Parasitol Res. 2002 Jun;88(6):489-94
pubmed: 12107469
Trop Biomed. 2014 Dec;31(4):579-91
pubmed: 25776583
Vet Res. 2018 Aug 2;49(1):79
pubmed: 30068382
Vet Res. 2020 Jun 15;51(1):78
pubmed: 32539772
PLoS One. 2013 Jun 28;8(6):e67570
pubmed: 23840742
Res Vet Sci. 2006 Dec;81(3):297-303
pubmed: 16516940
J Parasitol. 2003 Feb;89(1):92-8
pubmed: 12659309
Int J Biol Macromol. 2021 Dec 1;192:883-894
pubmed: 34656542
Parasitol Res. 2019 Jul;118(7):2247-2255
pubmed: 31081529
PLoS Negl Trop Dis. 2022 Nov 29;16(11):e0010929
pubmed: 36445875
Foodborne Pathog Dis. 2019 Nov;16(11):744-751
pubmed: 31215796
Exp Parasitol. 2016 Mar;162:35-42
pubmed: 26778819
Iran J Parasitol. 2022 Jul-Sep;17(3):375-384
pubmed: 36466014
Vet Res. 2018 Jul 13;49(1):59
pubmed: 30001738
Vet Res. 2021 Jan 7;52(1):6
pubmed: 33413587
PLoS Negl Trop Dis. 2022 Oct 31;16(10):e0010881
pubmed: 36315477
PLoS Negl Trop Dis. 2021 Oct 26;15(10):e0009865
pubmed: 34699522
Vet Res. 2020 Mar 14;51(1):43
pubmed: 32169101
PLoS Negl Trop Dis. 2020 Apr 27;14(4):e0008269
pubmed: 32339171
Korean J Parasitol. 2015 Feb;53(1):1-11
pubmed: 25748703
Parasit Vectors. 2018 Dec 5;11(1):625
pubmed: 30518426
Trop Biomed. 2020 Jun 1;37(2):458-470
pubmed: 33612815
Vet Res. 2020 Nov 23;51(1):139
pubmed: 33225967
Vet Res. 2019 Sep 23;50(1):70
pubmed: 31547875
Parasit Vectors. 2020 Feb 24;13(1):97
pubmed: 32093735
Exp Parasitol. 2018 Apr;187:59-66
pubmed: 29496524
Parasitol Int. 2016 Oct;65(5 Pt B):591-595
pubmed: 27495839
Vet Res. 2020 Sep 5;51(1):111
pubmed: 32891183
Parasitol Res. 2009 Jan;104(2):197-207
pubmed: 18987885
Exp Parasitol. 2022 Nov;242:108376
pubmed: 36089006
Res Vet Sci. 2020 Feb;128:1-8
pubmed: 31706217
Acta Trop. 2020 Nov;211:105592
pubmed: 32565198
Infect Immun. 1997 Nov;65(11):4806-12
pubmed: 9353069
Vet Res. 2022 Jun 23;53(1):48
pubmed: 35739604
Parasit Vectors. 2017 Sep 29;10(1):448
pubmed: 28962639
Vet Res. 2021 Aug 26;52(1):113
pubmed: 34446106
Parasite. 2015;22:37
pubmed: 26682577
Annu Rev Phytopathol. 2009;47:207-32
pubmed: 19400649
Vet Res. 2019 Dec 5;50(1):106
pubmed: 31806006
Vet Res. 2022 Oct 18;53(1):85
pubmed: 36258242
Parasitology. 2005 Apr;130(Pt 4):429-35
pubmed: 15830817
Acta Trop. 2015 Feb;142:57-63
pubmed: 25447831
Parasitol Res. 2016 Feb;115(2):615-22
pubmed: 26468148
Protein Pept Lett. 2019;26(3):170-175
pubmed: 30338728
Trop Biomed. 2021 Mar 1;38(1):160-171
pubmed: 33797541
Parasitol Today. 1998 Aug;14(8):318-23
pubmed: 17040798
Folia Parasitol (Praha). 2000;47(2):141-5
pubmed: 10945739
PLoS Negl Trop Dis. 2009;3(2):e377
pubmed: 19190745
Parasitol Res. 2015 May;114(5):2007-13
pubmed: 25758588
Vet Parasitol. 2021 Sep;297:109128
pubmed: 32402492
Mol Biochem Parasitol. 2004 Dec;138(2):165-70
pubmed: 15555728
J Parasitol. 2007 Apr;93(2):333-40
pubmed: 17539417
Vet Res. 2022 Mar 7;53(1):19
pubmed: 35255974
Acta Trop. 2021 Dec;224:106112
pubmed: 34453915
Vet Parasitol. 2016 Nov 15;231:43-46
pubmed: 27760716
Vet Parasitol. 2020 Sep;285:109234
pubmed: 32949838
Parasit Vectors. 2022 Dec 20;15(1):475
pubmed: 36539832
PLoS Negl Trop Dis. 2012;6(10):e1856
pubmed: 23071854
Vet Res. 2018 Dec 5;49(1):119
pubmed: 30518422