ABSTRACT
AIM: We investigated a proteome profile, protein-protein interaction and morphological changes of Mycobacterium tuberculosis after different times of eupomatenoid-5 (EUP-5) induction to evaluate the cellular response to the drug-induced damages. METHODS: The bacillus was induced to sub-minimal inhibitory concentration of EUP-5 at 12 h, 24 h and 48 h. The proteins were separated by 2D gel electrophoresis, identified by LC/MS-MS. Scanning electron microscopy and Search Tool for the Retrieval of Interacting Genes/Proteins analyses were performed. RESULTS: EUP-5 impacts mainly in M. tuberculosis proteins of intermediary metabolism and interactome suggests a multisite disturbance that contributes to bacilli death. Scanning electron microscopy revealed the loss of bacillary form. CONCLUSION: Some of the differentially expressed proteins have the potential to be drug targets such as citrate synthase (Rv0896), phosphoglycerate kinase (Rv1437), ketol-acid reductoisomerase (Rv3001c) and ATP synthase alpha chain (Rv1308).
Subject(s)
Benzofurans/pharmacology , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/metabolism , Phenols/pharmacology , Proteomics , Bacterial Proteins/drug effects , Bacterial Proteins/metabolism , Benzofurans/chemistry , Citrate (si)-Synthase/drug effects , Electrophoresis, Gel, Two-Dimensional , Genes, Bacterial/drug effects , Humans , Ketol-Acid Reductoisomerase/drug effects , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Mycobacterium tuberculosis/cytology , Mycobacterium tuberculosis/enzymology , Phenols/chemistry , Phosphoglycerate Kinase/drug effects , Protein Interaction Domains and Motifs , Proteome/analysis , Tandem Mass Spectrometry , Time Factors , Tuberculosis/drug therapy , Tuberculosis/microbiologyABSTRACT
AIM: To study the proteomic and morphological changes in Mycobacterium tuberculosis H37Rv exposed to subinhibitory concentration of isoniazid (INH). MATERIALS & METHODS: The bacillus was exposed to ½ MIC of INH at 12, 24 and 48 h. The samples' cells were submitted to scanning electron microscopy. The proteins were separated by 2D gel electrophoresis and identified by MS. RESULTS: INH exposure was able to alter the format, the multiplication and causing a cell swelling in the bacillus. The major altered proteins were related to the virulence, detoxification, adaptation, intermediary metabolism and lipid metabolism. CONCLUSION: The protein and morphological changes in M. tuberculosis induced by ½ MIC INH were related to defense mechanism of the bacillus or the action of INH therein.
