二硫键及其连接方式对防御素抗菌功能的影响研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2016.09.005

摘要/Abstract

摘要： 防御素是一类内源性、高度稳定、富含半胱氨酸的抗菌肽,对抗感染和宿主免疫调控具有重要作用。其序列内一般有6-8个保守半胱氨酸形成3-4对二硫键。二硫键对数和连接方式在维持防御素结构和抗菌功能方面具有重要作用,此外,高活性线型及低二硫键含量突变体在减少生产成本,简化生产流程方面具有重要作用。综述了防御素分子特征、结构和功能,在此基础上报道二硫键氧化还原状态、数量及其连接方式影响防御素抗菌活性的最新研究进展。

关键词: 防御素, 二硫键, 氧化还原, 连接, 抗菌活性

Abstract: Defensins are a class of endogenous,highly stable and cysteine-rich antimicrobial peptides,which play an important role in anti-microbial infection and the regulation of host immune. Six to eight conserved cysteines in the sequence form 3 to 4 pairs of disulfide bonds. The number of disulfide bond and its connection play an crucial role in maintaining the structure and antibacterial functions of defensins. In addition,mutants with high activity and low disulfide bond content also play an key role in reducing production costs and simplifying the production processes. The molecular characteristics,structure and function of defensins and their effects of redox status,number and connection mode of disulfide bonds on the antimicrobial activity are reviewed.

Key words: defensin, disulfide bond, oxidation and reduction, linkage, antimicrobial activity

陈惠娴, 毛若雨, 滕达, 王秀敏, 冯兴军, 王建华. 二硫键及其连接方式对防御素抗菌功能的影响研究进展[J]. 生物技术通报, 2016, 32(9): 32-37.

CHEN Hui-xian^{1, 2, 3}, MAO Ruo-yu^{1, 3}, TENG Da^{1, 3}, WANG Xiu-min^{1, 3}, FENG Xing-jun², WANG Jian-hua^{1, 3}. Review on Effects of Disulfide Bond and Its Connection on Antimicrobial Activity in Defensins[J]. Biotechnology Bulletin, 2016, 32(9): 32-37.

参考文献

[1] de Oliveira Dias R, Franco OL. Cysteine-stabilized αβ defensins：From a common fold to antibacterial activity[J]. Peptides, 2015, 72：64-72.
[2] Janeway Jr CA, Medzhitov R. Innate immune recognition[J]. Annual Review of Immunology, 2002, 20（1）：197-216.
[3] 付蓝宝, 于嘉林, 刘伟华. 防御素的生物学特性及其抗病基因工程[J]. 遗传, 2011, 33（5）：512-519.
[4] Selsted ME, Szklarek D, Lehrer RI. Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes[J]. Infection and Immunity, 1984, 45（1）：150-154.
[5] Lehrer RI, Ganz T, Szklarek D, et al. Modulation of the in vitro candidacidal activity of human neutrophil defensins by target cell metabolism and divalent cations[J]. Journal of Clinical Investigation, 1988, 81（6）：1829.
[6] Selsted ME, Harwig SS, Ganz T, et al. Primary structures of three human neutrophil defensins[J]. Journal of Clinical Investigation, 1985, 76（4）：1436.
[7] Daher KA, Selsted ME, Lehrer RI. Direct inactivation of viruses by human granulocyte defensins[J]. Journal of Virology, 1986, 60（3）：1068-1074.
[8] Lehrer RI, Daher K, Ganz T, et al. Direct inactivation of viruses by MCP-1 and MCP-2, natural peptide antibiotics from rabbit leukocytes[J]. Journal of Virology, 1985, 54（2）：467-472.
[9] Zhang Y, Lewis K. Fabatins：new antimicrobial plant peptides[J]. FEMS Microbiology Letters, 1997, 149（1）：59-64.
[10] Schibli DJ, Hunter HN, Aseyev V, et al. The solution structure the human beta-defensins lead to a better understanding of the potent bactericidal activity of HBD-3 against Sttaphylococcus aureus[J]. Journal of Biological Chemistry, 2002, 277（10）：8279-8289.
[11] Circo R, Skerlavaj B, Gennaro R, et al. Structural and functional characterization of hBD-1（Ser35）, a peptide deduced from a DFEB1 polymorphism[J]. Biochemical and Biophysical Research Communications, 2002, 293（1）：586-592.
[12] Gueguen Y, Herpin A, Aumelas A, et al. Characterization of a defensin from the oyster Crassostrea gigas recombinant production, folding, solution structure, antimicrobial activities, and gene expression[J]. Journal of Biological Chemistry, 2006, 281（1）：313-323.
[13] Smith JG, Nemerow GR. Mechanism of adenovirus neutralization by human α-defensins[J]. Cell Host & Microbe, 2008, 3（1）：11-19.
[14] 王少然, 杨雅麟, 张军, 等. 防御素构效关系研究进展[J]. 生物技术通报, 2011（4）：40-45.
[15] Ganz T. Defensins：antimicrobial peptides of innate immunity[J]. Nature Reviews Immunology, 2003, 3（9）：710-720.
[16] 王少然. 二硫键对菌丝霉素抗菌活性的影响及菌丝霉素多聚体在毕赤酵母中的表达[D]. 北京：中国农业科学院饲料研究所, 2011.
[17] 余兴邦, 郭锁链, 乌翠兰, 等. 防御素研究进展[J]. 动物医学进展, 2006, 27（8）：47-51.
[18] Yamada O, Sakamoto K, Tominaga M, et al. Cloning and heterologous expression of the antibiotic peptide（ABP）genes from Rhizopus oligosporus NBRC 8631[J]. Bioscience, Biotechnology, and Biochemistry, 2005, 69（3）：477-482.
[19] Ma D, Wang R, Liao W, et al. Identification and characterization of a novel antibacterial peptide, avian beta-defensin 2 from ducks[J]. Journal of Microbiology, 2009, 47（5）：610-618.
[20] 赵亚华, 徐来祥, 黄蓬亮, 等. 人类防御素的结构与功能研究进展[J]. 中国比较医学杂志, 2006, 16（7）：436-441.
[21] Chandrababu KB, Ho B, Yang D. Structure, dynamics, and activity of an all-cysteine mutated human β defensin-3 peptide analogue[J]. Biochemistry, 2009, 48（26）：6052-6061.
[22] Hoover DM, Wu Z, Tucker K, et al. Antimicrobial characterization of human β-defensin 3 derivatives[J]. Antimicrobial Agents And Chemotherapy, 2003, 47（9）：2804-2809.
[23] Landon C, Barbault F, Legrain M, et al. Rational design of peptides active against the gram positive bacteria Staphylococcus aureus[J]. Proteins：Structure, Function, and Bioinformatics, 2008, 72（1）：229-239.
[24] Varkey J, Singh S, Nagaraj R. Antibacterial activity of linear peptides spanning the carboxy-terminal β-sheet domain of arthropod defensins[J]. Peptides, 2006, 27（11）：2614-2623.
[25] Raj P, Antonyraj K, Karunakaran T. Large-scale synthesis and functional elements for the antimicrobial activity of defensins[J]. Biochem J, 2000, 347：633-641.
[26] Antcheva N, Morgera F, Creatti L, et al. Artificial beta-defensin based on a minimal defensin template[J]. Biochem J, 2009, 421：435-447.
[27] Zhang J, Yang Y, Teng D, et al. Expression of plectasin in Pichia pastoris and its characterization as a new antimicrobial peptide against Staphyloccocus and Streptococcus[J]. Protein Expression and Purification, 2011, 78（2）：189-196.
[28] Zhang Y, Teng D, Mao R, et al. High expression of a plectasin-deri-ved peptide NZ2114 in Pichia pastoris and its pharmacodynamics, postantibiotic and synergy against Staphylococcus aureus[J]. Applied Microbiology and Biotechnology, 2014, 98（2）：681-694.
[29] Zhang Y, Teng D, Wang X, et al. In vitro and in vivo characteriza-tion of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus[J]. Applied Microbiology and Biotechnology, 2015：99（15）：6255-6266.
[30] Yang Y, Teng D, Zhang J, et al. Characterization of recombinant plectasin：solubility, antimicrobial activity and factors that affect its activity[J]. Process Biochemistry, 2011, 46（5）：1050-1055.
[31] Cao X, Zhang Y, Mao R, et al. Design and recombination expression of a novel plectasin-derived peptide MP1106 and its properties against Staphylococcus aureus[J]. Applied Microbiology and Biotechnology, 2015, 99（6）：2649-2662.
[32] Peng Z, Wang A, Feng Q, et al. High-level expression, purification and characterisation of porcine β-defensin 2 in Pichia pastoris and its potential as a cost-efficient growth promoter in porcine feed[J]. Applied Microbiology And Biotechnology, 2014, 98（12）：5487-5497.
[33] Schroeder BO, Wu Z, Nuding S, et al. Reduction of disulphide bonds unmasks potent antimicrobial activity of human[bgr]-defensin 1[J]. Nature, 2011, 469（7330）：419-423.
[34] Maemoto A, Qu X, Rosengren KJ, et al. Functional analysis of the α-defensin disulfide array in mouse cryptdin-4[J]. Journal of Biological Chemistry, 2004, 279（42）：44188-44196.
[35] Wanniarachchi YA, Kaczmarek P, Wan A, et al. Human defensin 5 disulfide array mutants：disulfide bond deletion attenuates antibacterial activity against Staphylococcus aureus[J]. Biochemistry, 2011, 50（37）：8005-8017.
[36] Haag AF, Kerscher B, Dall’Angelo S, et al. Role of cysteine residues and disulfide bonds in the activity of a legume root nodule-specific, cysteine-rich peptide[J]. Journal of Biological Chemistry, 2012, 287（14）：10791-10798.
[37] Lee J, Lee D, Choi H, et al. Structure-activity relationships of the intramolecular disulfide bonds in coprisin, a defensin from the dung beetle[J]. BMB Reports, 2014, 47（11）：625.
[38] Chandrababu KB, Ho B, Yang D. Structure, dynamics, and activity of an all-cysteine mutated human β defensin-3 peptide analogue[J]. Biochemistry, 2009, 48（26）：6052-6061.
[39] Sharma H, Nagaraj R. Human β-Defensin 4 with non-native disulfide bridges exhibit antimicrobial activity[J]. PLoS One, 2015, 10（3）：e0119525.
[40] Valore EV, Ganz T. Laboratory production of antimicrobial peptides in native conformation[M]. Antibacterial Peptide Protocols. Humana Press, 1997：115-131.