title: Proteomic Analysis on Cercariae and Schistosomula in Reference to Potential Proteases Involved in Host Invasion of Schistosoma japonicum Larvae.
abstract: Schistosomiasis is a parasitic zoonosis posing great threat to human health. The infection is acquired by larval cercariae penetrating host skin and transforming into juveniles, schistosomula. Proteolytic enzymes secreted from the cercarial acetabular glands are known to aid to the skin penetration, but molecular mechanisms remain largely unclear. To profile the protein composition and identify potential invasive proteases, we developed a new method for simulating cercarial transformation and collecting schistosomula, and for the first time, we compared the proteomes of Schistosoma japonicum cercariae and schistosomula by using in-gel shotgun proteomic analysis. Totally, 1972 proteins were identified in association with ten main biological processes based on Gene Ontology analysis; 46 proteases were detected in cercariae, and among them, 25 proteases disappeared after penetrated. Notably, leishmanolysins and serine and cysteine proteases were found abundant but differentially expressed. Recombinant serine protease SjCE2b and cysteine protease SjCB2 were produced and used for validation of native proteins. Immunofluorescence and Western blotting assays detected SjCE2b and SjCB2 in cercariae but not in schistosomula, suggesting the two enzymes might be consumed upon skin migration. Our data comprehensively chart the proteomic changes during cercarial invasion, revealing the potential proteases involved, providing a platform for the development of molecular anti-infection strategy.
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Mu
authors.initials: M
authors.lastname: Liu
authors.affiliation: Key Laboratory of Parasite and Vector Biology of Ministry of Public Health, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention , 207 Rui Jin Er Road, Shanghai 200025, China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Ju
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Du
authors.affiliation: Key Laboratory of Parasite and Vector Biology of Ministry of Public Health, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention , 207 Rui Jin Er Road, Shanghai 200025, China.
authors.forename: Hai-Mo
authors.initials: HM
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Ji-Peng
authors.initials: JP
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Biochemistry, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Xu-Min
authors.initials: XM
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Parasite and Vector Biology of Ministry of Public Health, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention , 207 Rui Jin Er Road, Shanghai 200025, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.|Key Laboratory of Parasite and Vector Biology of Ministry of Public Health, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention , 207 Rui Jin Er Road, Shanghai 200025, China.|State Key Laboratory of Genetic Engineering, Department of Biochemistry, School of Life Sciences, Fudan University , 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D058487:Cercaria / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D002853:Chromatography, Liquid; D057056:Cysteine Proteases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D015801:Helminth Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008018:Life Cycle Stages / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification*; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000469:parasitology; D012908:Snails / Q000469:parasitology; D053719:Tandem Mass Spectrometry
chemical_list: D015801:Helminth Proteins; D010446:Peptide Fragments; D020543:Proteome; D011994:Recombinant Proteins; D057056:Cysteine Proteases; D012697:Serine Endopeptidases
keywords: Schistosoma japonicum; cercariae; cercarial transformation; invasion process; protease; proteomics; schistosomula