Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, from Coturnix coturnix | |
Feng, Feifei1,2; Chen, Chen3; Zhu, Wenjuan2; He, Weiyu1; Guang, Huijuan2; Li, Zheng2; Wang, Duo1; Liu, Jingze1; Chen, Ming5; Wang, Yipeng4; Yu, Haining1,2 | |
发表期刊 | FEBS JOURNAL |
ISSN | 1742-464X |
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 |
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 |
通讯作者 | Yu, HN, Hebei Normal Univ, Coll Life Sci, Shijiazhuang 050016, Hebei, Peoples R China.wyp010@163.com ; joannyu@live.cn |
作者部门 | 生物资源实验室 |
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. |
文章类型 | Article |
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] |
收录类别 | SCI |
语种 | 英语 |
关键词[WOS] | ANTIMICROBIAL PEPTIDE LL-37 ; INNATE IMMUNITY ; BOVINE NEUTROPHILS ; ANTIBACTERIAL PEPTIDES ; ONCORHYNCHUS-MYKISS ; SHEEP LEUKOCYTES ; RAINBOW-TROUT ; IDENTIFICATION ; BACTENECINS ; ANTIBIOTICS |
研究领域[WOS] | Biochemistry & Molecular Biology |
WOS记录号 | WOS:000289627800019 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.yic.ac.cn/handle/133337/4869 |
专题 | 海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室 |
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. |
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