Production of human antimicrobial peptide LL-37 in Escherichia coli using a thioredoxin-SUMO dual fusion system

被引:26
|
作者
Li, Yifeng [1 ]
机构
[1] Univ Texas Hlth Sci Ctr San Antonio, Dept Biochem, Prot Prod Core Facil, San Antonio, TX 78229 USA
关键词
Antimicrobial peptide; Dual-tag; Fusion protein; LL-37; SUMO; Thioredoxin; CATHELICIDIN LL-37; RECOMBINANT PRODUCTION; EXPRESSION; PURIFICATION; FAMILY; ROLES;
D O I
10.1016/j.pep.2012.10.008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
LL-37 is a human antimicrobial peptide that has been shown to possess multiple functions in host defense. In this report, the peptide was expressed as a fusion with a thioredoxin SUMO dual-tag. Upon SUMO protease mediated cleavage at the SUMO/peptide junction, LL-37 with its native N-terminus was generated. The released peptide was separated from the dual-tag and cleavage enzyme by size-exclusion chromatography. Mass spectrometry analysis proves that the recombinant peptide has a molecular weight as theoretically expected for its native form. The produced peptide displayed antimicrobial activity against Escherichia coli K-12. On average, 2.4 mg peptide was obtained from one liter of bacterial culture. Thus, the described approach provides an effective alternative for producing active recombinant LL-37 with its natural amino acid sequence in E. coli. (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:72 / 78
页数:7
相关论文
共 50 条
  • [1] Human antimicrobial peptide LL-37
    Yang, YH
    Ma, WR
    Zheng, GG
    Wu, KF
    PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS, 2003, 30 (06) : 847 - 851
  • [2] Antimicrobial peptide LL-37 in human atherosclerosis
    Edfeldt, K.
    Agerberth, B.
    Rottenberg, M. E.
    Gudmundsson, G. H.
    Mandal, K.
    Xu, Q. B.
    Hansson, G. K.
    Yan, Z. Q.
    ATHEROSCLEROSIS SUPPLEMENTS, 2006, 7 (03) : 407 - 407
  • [3] Rigidification of the Escherichia coli cytoplasm by the human antimicrobial peptide LL-37 revealed by superresolution fluorescence microscopy
    Zhu, Yanyu
    Mohapatra, Sonisilpa
    Weisshaar, James C.
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2019, 116 (03) : 1017 - 1026
  • [4] Lipopolysaccharide Phosphorylation by the WaaY Kinase Affects the Susceptibility of Escherichia coli to the Human Antimicrobial Peptide LL-37
    Bociek, Karol
    Ferluga, Sara
    Mardirossian, Mario
    Benincasa, Monica
    Tossi, Alessandro
    Gennaro, Renato
    Scocchi, Marco
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2015, 290 (32) : 19933 - 19941
  • [5] Real-time attack on single Escherichia coli cells by the human antimicrobial peptide LL-37
    Sochacki, Kem A.
    Barns, Kenneth J.
    Bucki, Robert
    Weisshaar, James C.
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (16) : E77 - E81
  • [6] Molecular farming in barley: production of human antimicrobial peptide LL-37
    Micchov, A.
    Holskov, E.
    Galuszka, P.
    Frbort, I.
    NEW BIOTECHNOLOGY, 2018, 44 : S27 - S27
  • [7] Involvement of the antimicrobial peptide LL-37 in human atherosclerosis
    Edfeldt, Kristina
    Agerberth, Birgitta
    Rottenberg, Martin E.
    Gudmundsson, Gudmundur H.
    Wang, Xiong-Biao
    Mandal, Kaushik
    Xu, Qingbo
    Yan, Zhong-qun
    ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2006, 26 (07) : 1551 - 1557
  • [8] Cloning, expression, isotope labeling, and purification of human antimicrobial peptide LL-37 in Escherichia coli for NMR studies
    Li, Yifeng
    Li, Xia
    Wang, Guangshun
    PROTEIN EXPRESSION AND PURIFICATION, 2006, 47 (02) : 498 - 505
  • [9] Expression of bloactive recombinant GSLL-39, a variant of human antimicrobial peptide LL-37, in Escherichia coli
    Yang, YH
    Zheng, GG
    Li, G
    Zhang, XJ
    Cao, ZY
    Rao, Q
    Wu, KF
    PROTEIN EXPRESSION AND PURIFICATION, 2004, 37 (01) : 229 - 235
  • [10] Recombinant production of human antimicrobial peptide LL-37 and its secondary structure
    Pavelka, Antonin
    Vacek, Lukas
    Norek, Adam
    Kobzova, Sarka
    Janda, Lubomir
    BIOLOGIA, 2024, 79 (01) : 263 - 273