Introduction: Infections by multidrug resistant Mycobacterium tuberculosis (MDR-TB) is a major public health challenge. Secretion of proteins by M. tuberculosis plays an important role in the pathogenesis of the bacterium. We compared the protein profiles of susceptible M. tuberculosis and MDR-TB isolates using proteomic analyses, namely two dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: The bacilli were cultured on Middlebrook 7H9 medium and bacterial colonies were mechanically disrupted and proteins were extracted by ammonium sulfate. The 2DE and MS analyses were performed using Ettan IP Gphor 3 isoelecteric system and Autoflex II TOF/TOF, respectively. Results: Our study showed that in comparison to the sensitive strains, 27 proteins were over-expressed in the MDR isolates and these proteins were mainly involved in the cellular metabolism, cell wall and membrane structures and bacterial respiration. Bactoferritin (Rv1876) has been shown to play a role in antibacterial resistance. Increased intensity of Rv2031c, a heat shock protein (Alpha-crystallin/HspX), was also observed in the whole cell lysate of the MDR-TB. This protein is a marker of the latent TB and has been proposed as a target for vaccine development. Conclusion: Our results identified proteins that are overexpressed in the resistant M. tuberculosis which could be used as antibacterial targets or vaccine candidates.
Introduction: Tuberculosis (TB) remains a deadly infectious disease despite all the efforts to reduce its incidence. Spread of multidrug resistant TB has seriously undermined the efforts to control the disease globally. In this study protein expression profile of MDR and sensitive isolates of MTB were analyzed and compared in order to identify proteins, which could be used in prevention, diagnosis and treatment. Methods: A sensitive and MDR isolate of Mycobacterium tuberculosis (MTB) were cultured on Middlebrook 7H9 medium and the whole cell lysates were subjected to native polyacrylamide gel electrophoresis (NPAGE) for protein expression profiling. Protein bands present in the MDR cell lysate that were not detected in the sensitive cell lysate were sent for identification by Matrixassisted laser desorption/ionization time-of-flightmass spectrometry (MALDI-TOF-MS). Results: Comparison of the protein expression profiles showed 6 bands that were not detected in the sensitive isolates. MTB Structural Annotation database search of the mass spectrometry results identified these bands as Rv3597c, Rv0379,Rv3614c, Rv0475, Rv0462, andRv0147and global transcriptional regulation, involvement in cell wall and cell processes and intermediary metabolism and respiration were the functions attributed to these proteins. Conclusion: Our results highlighted the complexities of linking protein expression to MDR phenotype as none of the proteins identified could be linked directly to drug resistance. The proteins identified in the present study were mostly those essential for survival or virulence of the bacteria, and could be used for diagnosis or as candidate vaccine, but with a better understanding of the function of these proteins their association with the MTB resistance to antibiotics might become clear.
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