Characterization of a c-type lysozyme of Scophthalmus maximus: Expression, activity, and antibacterial effect

doi:10.1016/j.fsi.2012.10.007

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	Characterization of a c-type lysozyme of Scophthalmus maximus: Expression, activity, and antibacterial effect
	Yu, Lan-ping 1,2; Sun, Bo-guang1 ; Li, Jun 1,3; Sun, Li 1; Sun, L
	2013
发表期刊	FISH & SHELLFISH IMMUNOLOGY
ISSN	1050-4648
卷号	34 期号:1 页码:46-54
文章类型	Article
摘要	Lysozyme is a key component of the innate immune system and plays an important role in antibacterial infection. In this study, we analyzed the expression and activity of a chicken-type (c-type) lysozyme (named SmLysC) from turbot (Scophthalmus maximus). SmLysC is composed of 143 residues and shares 67-90% overall sequence identities with the c-type lysozymes of a number of teleost fish. SmLysC possesses a typical c-type lysozyme domain, which contains the conserved residues E50 and D67 that form the putative catalytic site. SmLysC expression was detected, in increasing order, in head kidney, gill, heart, muscle, brain, spleen, blood, and liver. Bacterial infection caused significant inductions of SmLysC expression in head kidney, spleen, and liver in a time-dependent manner. Immunoblot analysis indicated that SmLysC has a subcellular localization in the extracellular milieu. Recombinant SmLysC (rSmLysC) was able to bind to bacterial cells and inhibit bacterial growth. Enzyme assay showed that the optimal temperature and pH of rSmLysC were 37 degrees C and pH 6.0 respectively. In contrast to rSmLysC, the mutant protein rSmLysCM1, which bears alanine substitutions at E50 and D67, displayed drastically reduced bacteriolytic activity. rSmLysC was able to inhibit the growth of several fish bacterial pathogens in a manner that depended on the dose of the protein; however, Gram-positive bacteria were in general more sensitive to rSmLysC than Gram-negative bacteria. Together these results indicate that SmLysC is a functional lysozyme that is likely to participate in innate immune defense against extracellular bacterial pathogens, in particular those of Gram-positive nature. (c) 2012 Elsevier Ltd. All rights reserved.; Lysozyme is a key component of the innate immune system and plays an important role in antibacterial infection. In this study, we analyzed the expression and activity of a chicken-type (c-type) lysozyme (named SmLysC) from turbot (Scophthalmus maximus). SmLysC is composed of 143 residues and shares 67-90% overall sequence identities with the c-type lysozymes of a number of teleost fish. SmLysC possesses a typical c-type lysozyme domain, which contains the conserved residues E50 and D67 that form the putative catalytic site. SmLysC expression was detected, in increasing order, in head kidney, gill, heart, muscle, brain, spleen, blood, and liver. Bacterial infection caused significant inductions of SmLysC expression in head kidney, spleen, and liver in a time-dependent manner. Immunoblot analysis indicated that SmLysC has a subcellular localization in the extracellular milieu. Recombinant SmLysC (rSmLysC) was able to bind to bacterial cells and inhibit bacterial growth. Enzyme assay showed that the optimal temperature and pH of rSmLysC were 37 degrees C and pH 6.0 respectively. In contrast to rSmLysC, the mutant protein rSmLysCM1, which bears alanine substitutions at E50 and D67, displayed drastically reduced bacteriolytic activity. rSmLysC was able to inhibit the growth of several fish bacterial pathogens in a manner that depended on the dose of the protein; however, Gram-positive bacteria were in general more sensitive to rSmLysC than Gram-negative bacteria. Together these results indicate that SmLysC is a functional lysozyme that is likely to participate in innate immune defense against extracellular bacterial pathogens, in particular those of Gram-positive nature. (c) 2012 Elsevier Ltd. All rights reserved.
关键词	C-type Lysozyme Scophthalmus Maximus Bacteriolytic Catalytic Site
学科领域	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI	10.1016/j.fsi.2012.10.007
URL	查看原文
收录类别	SCI
语种	英语
WOS研究方向	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS类目	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS记录号	WOS:000314743600006
WOS关键词	GOOSE-TYPE LYSOZYME ; MOLECULAR CHARACTERIZATION ; IMMUNOPROTECTIVE ANALYSIS ; VACCINE CANDIDATE ; CDNA ; IDENTIFICATION ; PHYLOGENY ; INFECTION ; PATTERN ; ANTIGEN
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
引用统计	被引频次：48[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/16538
专题	实验海洋生物学重点实验室
通讯作者	Sun, L
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Lake Super State Univ, Sch Biol Sci, Sault Ste Marie, MI USA
第一作者单位	实验海洋生物学重点实验室
推荐引用方式 GB/T 7714	Yu, Lan-ping,Sun, Bo-guang,Li, Jun,et al. Characterization of a c-type lysozyme of Scophthalmus maximus: Expression, activity, and antibacterial effect[J]. FISH & SHELLFISH IMMUNOLOGY,2013,34(1):46-54.
APA	Yu, Lan-ping,Sun, Bo-guang,Li, Jun,Sun, Li,&Sun, L.(2013).Characterization of a c-type lysozyme of Scophthalmus maximus: Expression, activity, and antibacterial effect.FISH & SHELLFISH IMMUNOLOGY,34(1),46-54.
MLA	Yu, Lan-ping,et al."Characterization of a c-type lysozyme of Scophthalmus maximus: Expression, activity, and antibacterial effect".FISH & SHELLFISH IMMUNOLOGY 34.1(2013):46-54.

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