| Autor: |
Han X; School of Life Science, Liaoning Normal University, Dalian, China., Kou Z; School of Life Science, Liaoning Normal University, Dalian, China., Jiang F; Department of Clinical Laboratory, the First Affiliated Hospital of Dalian Medical University, Dalian, China., Sun X; School of Life Science, Liaoning Normal University, Dalian, China., Shang D; School of Life Science, Liaoning Normal University, Dalian, China.; Liaoning Provincial Key Laboratory of Biotechnology and Drug Discovery, Liaoning Normal University, Dalian, China. |
| Abstrakt: |
To investigate the intracellular mechanisms of seven Trp-containing peptides in clinically isolated multidrug-resistant Pseudomonas aeruginosa (MRPA0108). The results showed that the Trp-containing peptides had high antibacterial activity against the MRPA0108 strain, with minimal inhibitory concentration (MIC) values ranging from 6.25 to 25 μM. The peptides rapidly and completely killed the MRPA0108 at a concentration of 16 × MIC at 60-90 min. The Trp-containing peptides were found to penetrate the bacterial cell membrane and accumulate in the cells. A DNA gel retardation assay indicated that the peptides were able to bind with the genomic DNA of MRPA0108 cells; L5W exhibited a stronger DNA binding ability than that of the other peptides, and the ratio of peptide to DNA was 0.62/1. Next, the UV absorption spectrum of the DNA indicated that L5W interacted with the MRPA0108 genomic DNA and intercalated into the groove of the DNA molecule, resulting in loosening of the double-helical structure of the originally contracted DNA and leading to the occurrence of a hyperchromic effect. The circular dichroism spectrum suggested that I1W and L5W associated with the DNA via a trench combination mode resulting from the compact structure of the DNA double helix and reduction in ππ accumulation between base pairs. Furthermore, real-time quantitative PCR demonstrated that the Trp-containing peptides could downregulate the expression of DNA replication-initiating genes in MRPA0108 cells. MRPA0108 DNA may be a potential active target for the antimicrobial activity of Trp-containing peptides. |
