碱性氨基酸对螺旋型抗菌肽生物活性的影响

doi:10.7503/cjcu20170596

高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (4): 681.doi: 10.7503/cjcu20170596

碱性氨基酸对螺旋型抗菌肽生物活性的影响

赵文采, 韩丽丽, 彭颖君, 王晓静, 刘晟宇, 李鹏飞, 黄宜兵(), 陈育新

吉林大学分子酶学工程教育部重点实验室, 生命科学学院, 长春 130021

收稿日期:2017-09-01 出版日期:2018-04-10 发布日期:2018-03-26
作者简介:联系人简介: 黄宜兵, 男, 博士, 副教授, 主要从事多肽药物方面的研究. E-mail:huangyibing@jlu.edu.cn
基金资助:
国家自然科学基金(批准号: 81373455)、吉林省自然科学基金(批准号: 20180101250JC)和超分子结构与材料国家重点实验室开放课题(批准号: SKLSSM201708)资助

Effect of Basic Amino Acids on the Biological Activity of Helical Antimicrobial Peptide^†

ZHAO Wencai, HAN Lili, PENG Yingjun, WANG Xiaojing, LIU Shengyu, LI Pengfei, HUANG Yibing^*(), CHEN Yuxin

Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education,School of Life Science, Jilin University, Changchun 130012, China

Received:2017-09-01 Online:2018-04-10 Published:2018-03-26
Contact: HUANG Yibing E-mail:huangyibing@jlu.edu.cn
Supported by:
† Supported by the National Natural Science Foundation of China(No.81373445), the Natural Science Foundation of Jilin Province, China(No.20180101250JC) and the Open Project of State Key Laboratory of Supramolecular Structure and Materials, China(No.SKLSSM201708)

摘要/Abstract

摘要：

以螺旋型抗菌肽HPRP-A1为模板, 利用精氨酸(R)对HPRP-A1序列中的赖氨酸(K)进行不同位点、不同数量的取代, 结合改造前后抗菌肽的螺旋度、疏水性及自聚集常数等理化性质, 研究了螺旋型抗菌肽结构与活性的相关性. 进一步结合脂质体模拟和细胞膜穿透实验, 对螺旋型抗菌肽与不同类型细胞膜的相互作用过程进行了研究. 结果表明, 增加精氨酸的取代数量, 会增强抗菌肽的疏水性和螺旋度, 导致螺旋型抗菌肽对真核细胞的毒性增强; 但精氨酸的增加会伴随着抗菌肽自聚集能力的降低以及抗菌肽对细菌细胞膜的渗透性降低, 导致抗菌肽的抗菌活性降低. 本研究对于设计和改造具有应用前景的螺旋型抗菌肽具有重要指导意义.

关键词: 抗菌肽, 精氨酸, 赖氨酸, 脂质体模拟膜

Abstract:

In order to investigate the effect of basic amino acids on the biological activity of antimicrobial peptides, an α helical antimicrobial peptides of HPRP-A1 was used to design several peptide derivatives with different K/R ratio by substitution lysine(K) with arginine(R). The relationship between the structure and activity of helical antimicrobial peptides was studied by combining the biophysical properties such as helicity, hydrophobicity and peptide association parameter of antimicrobial peptides. The interaction between helical antimicrobial peptides and different types of cell membranes was further studied by liposome mimic membrane and the cell permeation ability of peptide was examined by 1-N-pbenyl-naphtylamine(NPN) and o-nitrophenyl-β-D-galactopyranoside(ONPG). The results confirmed that the increasing numbers of arginine substitution increased the hydrophobicity and helicity of the antimicrobial peptides, resulting in increasing toxicity of the helical antimicrobial peptides against eukaryotic cell membrane. However, the increasing of arginine was accompanied by decreasing in the ability of peptide association and the permeability of the antimicrobial peptide against the prokaryotic cell membrane, resulting in decreasing of the antibacterial activity of the antimicrobial peptide.

Key words: Antimicrobial peptide, Arginin, Lysine, Liposome mimic membrane

中图分类号:

O629.7

TrendMD:

赵文采, 韩丽丽, 彭颖君, 王晓静, 刘晟宇, 李鹏飞, 黄宜兵, 陈育新. 碱性氨基酸对螺旋型抗菌肽生物活性的影响. 高等学校化学学报, 2018, 39(4): 681.

ZHAO Wencai, HAN Lili, PENG Yingjun, WANG Xiaojing, LIU Shengyu, LI Pengfei, HUANG Yibing, CHEN Yuxin. Effect of Basic Amino Acids on the Biological Activity of Helical Antimicrobial Peptide^†. Chem. J. Chinese Universities, 2018, 39(4): 681.

图/表 6

Table 1 Biophysical properties of HPRP-A1 and its derivatives*

Peptide	Amino acid sequence	M_w	Hydrophobicity t_R0/min	P_A
HPRP-A1	Ac-FKKLKKLFSKLWNWK-amide	2036.54	46.93	2.342
HPRP-A1-2R	Ac-FRKLKKLFSKLWNWR-amide	2092.56	47.28	2.205
HPRP-A1-4R	Ac-FRKLKRLFSRLWNWR-amide	2148.59	47.65	1.229
HPRP-A1-6R	Ac-FRRLRRLFSRLWNWR-amide	2204.62	48.45	1.092

Fig.1 CD spectra of HPRP-A1 and its derivatives(A) In the presence of PBS buffer and 2,2,2-Trifluoroethanol(TFE)(volume ratio 1:1);(B) in PBS buffer(50 mmol/L KH2PO4/K2HPO4+100 mmol/L KCl, pH=7.4) at pH=7.4, 25 ℃(B).

Table 2 Biological activities of HPRP-A1 and its derivatives*

Peptide	MIC/(μmol·L^-1)				MHC/(μmol·L^-1)	Therapeutic index
Peptide	P. aeruginosa	S. aereus	E. coil	B. subtilis	MHC/(μmol·L^-1)	Therapeutic index
HPRP-A1	16	16	8	8	64	5.33
HPRP-A1-2R	32	32	8	4	64	3.37
HPRP-A1-4R	64	16	16	4	32	1.28
HPRP-A1-6R	64	32	32	8	32	0.94

Fig.2 Outer membrane permeabilization assay of HPRP-A1 and its derivativesNPN uptake in P. aeruginosa ATCC 27853 was continuously measured by fluorescence spectrophotometer for 12 min at 25 ℃ endued by 4 μmol/L HPRP-A1 and its derivatives(A) and 2, 4 and 8 μmol/L HPRP-A1(B).

Fig.3 Inner membrane permeabilization assay of HPRP-A1 and its derivativesRelease of cytoplasmic β-galactosidase activity were measured by UV-visible spectrophotometer at 420 nm after adding ONPG(30 mmol/L) and various concentrations peptides into E. coli ML-35 bacterial suspension.

Fig.4 Fluorescence emission spectra(A, B) and Stern-Volmer plot(C, D) of peptides with various liposome models at 25 ℃Stern-Volmer plots were obtained by the sequential addition of the fluorescence quencher KI. PC/PG liposomes(mass fraction 7:3) were used to mimic prokaryotic membrane in (A, C) and PC/cholesterol(mass fraction 8:1) liposomes were used to mimic eukaryotic membrane in (B, D), respectively.

参考文献 27

[1]	Zhang Y. H., Li L. C., Yuan W. C., Zhang X. M., Chem. Res. Chinese Universities, 2015, 31(3), 381-387
[2]	Feng H. Y., Gao L., Ye X. H., Wang L., Xue Z. C., Kong J. M., Li L. Z., Chem. Res. Chinese Universities, 2017, 33(1), 155-159
[3]	Cheng S. Q., Liang G. D., Jiang X. F., Wang C., Liu K. L., Chem. J. Chinese Universities, 2016, 37(7), 1287-1292
	(程思绮, 梁国栋, 姜喜凤, 王潮, 刘克良. 高等学校化学学报, 2016, 37(7), 1287-1292)
[4]	Andersson D. I., Hughes D., Kubicek-Sutherland J. Z., Drug Resist. Updates, 2016, 26, 43-57
[5]	Hancock R. E. W., Sahl H. G., Nat. Biotechnol., 2006, 24, 1551-1557
[6]	Huang Y. B., Zhai N. C., Gao G., Chen Y. X., Chem. J. Chinese Universities, 2012, 33(6), 1252-1258
	(黄宜兵, 翟乃翠, 高贵, 陈育新. 高等学校化学学报, 2012, 33(6), 1252-1258)
[7]	Huang Y. B., He L. Y., Li G. R., Zhai N. C., Jiang H. Y., Chen Y. X., Protein & Cell, 2014, 5, 631-642
[8]	Huang Y. B., Huang J. F., Chen Y. X., Protein & Cell, 2010, 1, 143-152
[9]	Bechara C., Sagan S., Febs Lett., 2013, 587, 1693-1702
[10]	Vasconcelos L., Parn K., Langel U., Ther. Deliv., 2013, 4, 573-591
[11]	Hao X. Y., Yan Q. Y., Zhao J., Wang W. R., Huang Y. B., Chen Y. X., PLoS One, 2015, 10, e0138911
[12]	Schmidt N. W., Tai K. P., Kamdar K., Mishra A., Lai G. H., Zhao K., Ouellette A. J., Wong G. C., J. Biol. Chem., 2012, 287, 21866-21872
[13]	Zou G. Z., de Leeuw E., Li C., Pazgier M., Li C. Q., Zeng P., Lu W. Y., Lubkowski J., Lu W., J. Biol. Chem., 2007, 282, 19653-19665
[14]	Huang Y. B., Wang X. F., Wang H. Y., Liu Y., Chen Y. X., Mol. Cancer Ther., 2011, 10, 416-426
[15]	Mant C. T., Chen Y., Hodges R. S., J. Chromatogr. A, 2003, 1009, 29-43
[16]	Chen Y. X., Guarnieri M. T., Vasil A. I., Vasil M. L., Mant C. T., Hodges R. S., Antimicrob. Agents Chemother., 2007, 51, 1398-1406
[17]	Yi T. H., Huang Y. B., Chen Y. X., Journal of Peptide Science, 2015, 21, 46-52
[18]	Sun S. Y., Zhao G. X., Huang Y. B, Cai M. J., Shan Y. P., Wang H. D., Chen Y. X., Sci. Rep., 2016, 6, e29145
[19]	Chen Y. X., Mant C. T., Farmer S. W., Hancock R. E. W., Vasil M. L., Hodges R. S., J. Biol. Chem., 2005, 280, 12316-12329
[20]	Huang H. W., Biochemistry, 2000, 39, 8347-8352
[21]	Yi T. H., Huang Y. B., Chen Y. X., Chem. Bio. Drug Des., 2015, 85, 598-607
[22]	Eriksson M., Nielsen P. E., Good L., J. Biol. Chem., 2002, 277, 7144-7147
[23]	Li L. B., Vorobyov I., Allen T. W., J. Phys. Chem. B, 2013, 117, 11906-11920
[24]	Harris F., Dennison S. R., Singh J., Phoenix D. A., Med. Res. Rev., 2013, 33, 190-234
[25]	Lopez-Exposito I., Amigo L., Recio I., BBA-Biomembranes,2008, 1778, 2444-2449
[26]	Mant C. T., Kovacs J. M., Kim H. M., Pollock D. D., Hodges R. S., Biopolymers,2009, 92, 573-595
[27]	Qiao Y., Gao Z. H., Liu Y., Cheng Y., Yu M. X., Duan Y., Liu Y., Biomed. Res. Int., 2014, 2014, e869186

碱性氨基酸对螺旋型抗菌肽生物活性的影响

Effect of Basic Amino Acids on the Biological Activity of Helical Antimicrobial Peptide^†

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 27

相关文章 15

编辑推荐

Metrics

本文评价

[1]	郑海娇, 姜丽艳, 贾琼. 精氨酸功能化磁性纳米材料的制备及在磷酸化肽富集中的应用[J]. 高等学校化学学报, 2021, 42(3): 717.
[2]	王煜瑶, 张强, 于吉红. 多级孔NaX分子筛的合成及CO₂吸附性能[J]. 高等学校化学学报, 2020, 41(4): 616.
[3]	常俊朋, 赵佳瑞, 陈思佳, 孟凯, 石微妮, 李瑞芳. 抗菌肽SAMP1及其类似肽的构效关系[J]. 高等学校化学学报, 2019, 40(4): 705.
[4]	赵麒, 何婉莹, 段莉洁, 张瑜, 于双江, 高光辉. 基于席夫碱键的可注射糖肽水凝胶的制备及性能[J]. 高等学校化学学报, 2016, 37(9): 1750.
[5]	李蕾, 黄翠英, 姜笑楠, 高希婵, 王长生. 精氨酸侧链和核酸碱基间离子氢键作用强度分析[J]. 高等学校化学学报, 2016, 37(8): 1460.
[6]	黄义玲, 高雪峰. L-缬氨酸对精氨酸酶Ⅰ抑制作用的分子动力学模拟[J]. 高等学校化学学报, 2016, 37(5): 928.
[7]	安会琴, 于育才, 闫琳, 吴婷婷, 李晓峰, 贺晓凌, 赵莉芝, 黄唯平. 赖氨酸辅助高分散金粒子修饰氮掺杂TiO₂纳米管催化剂的合成及光催化性能[J]. 高等学校化学学报, 2016, 37(11): 2034.
[8]	张栋梁, 黄宇彬, 李晓媛. 基于邻苯二酚的可降解黏合剂的制备及表征[J]. 高等学校化学学报, 2015, 36(6): 1236.
[9]	于倩, 何颖芳, 陈媛, 杨文江, 张春, 陆洁. ^99mTc标记蝶呤-赖氨酸衍生物的制备及生物性能评价[J]. 高等学校化学学报, 2015, 36(12): 2446.
[10]	王梅, 王军, 步宇翔. 与蛋白质调控DNA空穴迁移相关的具有负离解能特征的亚稳态氢键[J]. 高等学校化学学报, 2015, 36(11): 2271.
[11]	于岚岚, 毛烨炫, 白希希, 冉瑜, 李爱荣, 朱艳艳, 于斐, 屈凌波. 具有双活性序列的新型抗菌肽的设计及性质[J]. 高等学校化学学报, 2013, 34(5): 1166.
[12]	赵志强, 邱辉华, 姚军, 陈永峰. 基于化学衍生和CID碎裂模式鉴定蛋白精氨酸-ADP-核糖基化的新方法[J]. 高等学校化学学报, 2013, 34(12): 2704.
[13]	梁亚琴, 胡志勇, 曹端林, 梁栋. 手性赖氨酸基Gemini表面活性剂的表面性能[J]. 高等学校化学学报, 2013, 34(12): 2783.
[14]	张建成, 丁建勋, 肖春生, 贺超良, 庄秀丽, 杨亚楠, 陈学思. 肿瘤酸度响应性聚(L-赖氨酸)-阿霉素键合药的制备与表征[J]. 高等学校化学学报, 2012, 33(12): 2809.
[15]	于岚岚, 冉瑜, 白希希, 李爱荣, 朱艳艳, 覃韵, 屈凌波. 新型抗菌肽的设计、活性研究及与磷脂相互作用的计算模拟[J]. 高等学校化学学报, 2012, 33(12): 2681.

碱性氨基酸对螺旋型抗菌肽生物活性的影响

Effect of Basic Amino Acids on the Biological Activity of Helical Antimicrobial Peptide†

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 27

相关文章 15

编辑推荐

Metrics

本文评价

Effect of Basic Amino Acids on the Biological Activity of Helical Antimicrobial Peptide^†