PMID- 30894414 OWN - NLM STAT- MEDLINE DCOM- 20191209 LR - 20210205 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 294 IP - 19 DP - 2019 May 10 TI - A small mycobacteriophage-derived peptide and its improved isomer restrict mycobacterial infection via dual mycobactericidal-immunoregulatory activities. PG - 7615-7631 LID - 10.1074/jbc.RA118.006968 [doi] AB - Mycobacteriophages express various peptides/proteins to infect Mycobacterium tuberculosis (M. tb). Particular attention has been paid to mycobacteriophage-derived endolysin proteins. We herein characterized a small mycobacteriophage-derived peptide designated AK15 with potent anti-M. tb activity. AK15 adopted cationic amphiphilic alpha-helical structure, and on the basis of this structure, we designed six isomers with increased hydrophobic moment by rearranging amino acid residues of the helix. We found that one of these isomers, AK15-6, exhibits enhanced anti-mycobacterial efficiency. Both AK15 and AK15-6 directly inhibited M. tb by trehalose 6,6'-dimycolate (TDM) binding and membrane disruption. They both exhibited bactericidal activity, cell selectivity, and synergistic effects with rifampicin, and neither induced drug resistance to M. tb They efficiently attenuated mycobacterial load in the lungs of M. tb-infected mice. We observed that lysine, arginine, tryptophan, and an alpha-helix are key structural requirements for their direct anti-mycobacterial action. Of note, they also exhibited immunomodulatory effects, including inhibition of proinflammatory response in TDM-stimulated or M. tb-infected murine bone marrow-derived macrophages (BMDMs) and M.tb-infected mice and induction of only a modest level of cytokine (tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6)) production in murine BMDMs and a T-cell cytokine (interferin-gamma (IFN-gamma) and TNF-alpha) response in murine lung and spleen. In summary, characterization of a small mycobacteriophage-derived peptide and its improved isomer revealed that both efficiently restrain M. tb infection via dual mycobactericidal-immunoregulatory activities. Our work provides clues for identifying small mycobacteriophage-derived anti-mycobacterial peptides and improving those that have cationic amphiphilic alpha-helices. CI - (c) 2019 Yang et al. FAU - Yang, Yang AU - Yang Y AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu. FAU - Liu, Zhen AU - Liu Z AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu. FAU - He, Xiaoqin AU - He X AD - the Key Laboratory of Animal Models and Human Disease Mechanisms, Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Kunming 650223, Yunnan. AD - the National Health Commission Key Laboratory of Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, Jiangsu Province. FAU - Yang, Juanjuan AU - Yang J AD - the Institute of Pharmaceutical Biotechnology and Engineering, College of Biological Science and Technology, Fuzhou University, Fuzhou 350108, and. FAU - Wu, Jing AU - Wu J AD - the School of Basic Medical Sciences, Kunming Medical University, Kunming 650500, Yunnan, China. FAU - Yang, Hailong AU - Yang H AD - the School of Basic Medical Sciences, Kunming Medical University, Kunming 650500, Yunnan, China. FAU - Li, Min AU - Li M AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu. FAU - Qian, Qian AU - Qian Q AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu. FAU - Lai, Ren AU - Lai R AD - the Key Laboratory of Animal Models and Human Disease Mechanisms, Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Kunming 650223, Yunnan, rlai@mail.kiz.ac.cn. FAU - Xu, Wei AU - Xu W AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu, xuweifd828@126.com. FAU - Wei, Lin AU - Wei L AUID- ORCID: 0000-0003-3359-2471 AD - From the Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, Jiangsu, weilin1005@126.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190320 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (Anti-Bacterial Agents) RN - 0 (Antimicrobial Cationic Peptides) RN - 0 (Viral Proteins) RN - VJT6J7R4TR (Rifampin) SB - IM MH - Animals MH - Anti-Bacterial Agents/chemistry/*pharmacology MH - Antimicrobial Cationic Peptides/agonists/chemistry/*pharmacology MH - Drug Synergism MH - Humans MH - Macrophages/immunology/pathology MH - Mice MH - Mice, Inbred BALB C MH - Mycobacteriophages/*chemistry MH - Mycobacterium tuberculosis/*immunology/virology MH - Rifampin/agonists/pharmacology MH - Tuberculosis, Pulmonary/*drug therapy/immunology/pathology MH - Viral Proteins/chemistry/*pharmacology PMC - PMC6514635 OTO - NOTNLM OT - Mycobacterium tuberculosis OT - amino acid OT - anti-mycobacterial peptide OT - antibiotic OT - antimicrobial peptide (AMP) OT - drug action OT - drug resistance OT - endolysin OT - hydrophobic moment OT - immune regulation OT - mycobacteriophage OT - peptides OT - structure-function OT - alpha-helix COIS- The authors declare that they have no conflicts of interest with the contents of this article EDAT- 2019/03/22 06:00 MHDA- 2019/12/18 06:00 PMCR- 2020/05/10 CRDT- 2019/03/22 06:00 PHST- 2018/12/07 00:00 [received] PHST- 2019/03/07 00:00 [revised] PHST- 2019/03/22 06:00 [pubmed] PHST- 2019/12/18 06:00 [medline] PHST- 2019/03/22 06:00 [entrez] PHST- 2020/05/10 00:00 [pmc-release] AID - S0021-9258(20)35459-4 [pii] AID - RA118.006968 [pii] AID - 10.1074/jbc.RA118.006968 [doi] PST - ppublish SO - J Biol Chem. 2019 May 10;294(19):7615-7631. doi: 10.1074/jbc.RA118.006968. Epub 2019 Mar 20.