Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix

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	Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix
	Feng, Feifei 1,2; Chen, Chen 3; Zhu, Wenjuan 2; He, Weiyu 1; Guang, Huijuan 2; Li, Zheng 2; Wang, Duo 1; Liu, Jingze 1; Chen, Ming 5; Wang, Yipeng4
刊名	FEBS JOURNAL
	2011-05-01
卷号	278 期号:9 页码:1573-1584
关键词	Cathelicidin Coturnix Coturnix Expression Molecular Cloning Structure And Function
ISSN号	1742-464X
通讯作者	Yu, HN, Hebei Normal Univ, Coll Life Sci, Shijiazhuang 050016, Hebei, Peoples R China.wyp010@163.com ; joannyu@live.cn
产权排序	[Feng, Feifei; He, Weiyu; Wang, Duo; Liu, Jingze; Yu, Haining] Hebei Normal Univ, Coll Life Sci, Shijiazhuang 050016, Hebei, Peoples R China; [Feng, Feifei; Zhu, Wenjuan; Guang, Huijuan; Li, Zheng; Yu, Haining] Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian, Liaoning, Peoples R China; [Chen, Chen] Shaanxi Univ Technol, Coll Biol Sci & Engn, Hanzhong, Shaanxi, Peoples R China; [Wang, Yipeng] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Biol Resources Lab, Yantai 264003, Shandong, Peoples R China; [Chen, Ming] Chengdu Univ Tradit Chinese Med, Teaching Hosp, Dept Nephrol, Chengdu, Peoples R China
文献子类	Article
英文摘要	Cathelicidins comprise a family of antimicrobial peptides sharing a highly conserved cathelin domain, which play a central role in the early innate host defense against infection. In the present study, we report three novel avian cathelicidin orthologs cloned from a constructed spleen cDNA library of Coturnix coturnix, using a nested-PCR-based cloning strategy. Three coding sequences containing ORFs of 447, 465 and 456 bp encode three mature antimicrobial peptides (named Cc-CATH1, 2 and 3) of 26, 32 and 29 amino acid residues, respectively. Phylogenetic analysis indicated that precursors of Cc-CATHs are significantly conserved with known avian cathelicidins. Synthetic Cc-CATH2 and 3 displayed broad and potent antimicrobial activity against most of the 41 strains of bacteria and fungi tested, especially the clinically isolated drug-resistant strains, with minimum inhibitory concentration values in the range 0.3-2.5 mu m for most strains with or without the presence of 100 mm NaCl. Cc-CATH2 and 3 showed considerable reduction of cytotoxic activity compared to other avian cathelicidins, with average IC50 values of 20.18 and 17.16 mu m, respectively. They also exerted a negligible hemolytic activity against human erythrocytes, lysing only 3.6% of erythrocytes at a dose up to 100 mu g center dot mL-1. As expected, the recombinant Cc-CATH2 (rCc-CATH2) also showed potent bactericidal activity. All these features of Cc-CATHs encourage further studies aiming to estimate their therapeutic potential as drug leads, as well as coping with current widespread antibiotic resistance, especially the new prevalent and dangerous 'superbug' that is resistant to almost all antibiotics.; Cathelicidins comprise a family of antimicrobial peptides sharing a highly conserved cathelin domain, which play a central role in the early innate host defense against infection. In the present study, we report three novel avian cathelicidin orthologs cloned from a constructed spleen cDNA library of Coturnix coturnix, using a nested-PCR-based cloning strategy. Three coding sequences containing ORFs of 447, 465 and 456 bp encode three mature antimicrobial peptides (named Cc-CATH1, 2 and 3) of 26, 32 and 29 amino acid residues, respectively. Phylogenetic analysis indicated that precursors of Cc-CATHs are significantly conserved with known avian cathelicidins. Synthetic Cc-CATH2 and 3 displayed broad and potent antimicrobial activity against most of the 41 strains of bacteria and fungi tested, especially the clinically isolated drug-resistant strains, with minimum inhibitory concentration values in the range 0.3-2.5 mu m for most strains with or without the presence of 100 mm NaCl. Cc-CATH2 and 3 showed considerable reduction of cytotoxic activity compared to other avian cathelicidins, with average IC(50) values of 20.18 and 17.16 mu m, respectively. They also exerted a negligible hemolytic activity against human erythrocytes, lysing only 3.6% of erythrocytes at a dose up to 100 mu g center dot mL-1. As expected, the recombinant Cc-CATH2 (rCc-CATH2) also showed potent bactericidal activity. All these features of Cc-CATHs encourage further studies aiming to estimate their therapeutic potential as drug leads, as well as coping with current widespread antibiotic resistance, especially the new prevalent and dangerous 'superbug' that is resistant to almost all antibiotics.
学科主题	Biochemistry & Molecular Biology
WOS关键词	ANTIMICROBIAL PEPTIDE LL-37 ; INNATE IMMUNITY ; BOVINE NEUTROPHILS ; ANTIBACTERIAL PEPTIDES ; ONCORHYNCHUS-MYKISS ; SHEEP LEUKOCYTES ; RAINBOW-TROUT ; IDENTIFICATION ; BACTENECINS ; ANTIBIOTICS
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
WOS记录号	WOS:000289627800019
资助机构	National High-Tech R D Project [2007AA09Z403]; National Natural Science Foundation of China [30530080]; Department of Science and Technology of Shandong Province [2006GG2205023]; Chinese Academy of Sciences for Key Topics in Innovation Engineering [KZCX2-YW-QN209]
公开日期	2011-07-22
内容类型	期刊论文
源URL	[http://ir.yic.ac.cn/handle/133337/4870]
专题	烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室
作者单位	1.Hebei Normal Univ, Coll Life Sci, Shijiazhuang 050016, Hebei, Peoples R China 2.Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian, Liaoning, Peoples R China 3.Shaanxi Univ Technol, Coll Biol Sci & Engn, Hanzhong, Shaanxi, Peoples R China 4.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Biol Resources Lab, Yantai 264003, Shandong, Peoples R China 5.Chengdu Univ Tradit Chinese Med, Teaching Hosp, Dept Nephrol, Chengdu, Peoples R China
推荐引用方式 GB/T 7714	Feng, Feifei,Chen, Chen,Zhu, Wenjuan,et al. Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix[J]. FEBS JOURNAL,2011,278(9):1573-1584.
APA	Feng, Feifei.,Chen, Chen.,Zhu, Wenjuan.,He, Weiyu.,Guang, Huijuan.,...&Yu, Haining.(2011).Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix.FEBS JOURNAL,278(9),1573-1584.
MLA	Feng, Feifei,et al."Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix".FEBS JOURNAL 278.9(2011):1573-1584.