Proteomic analysis of streptomycin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

Abstract

BackgroundStreptomycin (SM) is a broad spectrum antibiotic and is an important component of any anti-tuberculosis therapy regimen. Several mechanisms have been proposed to explain the emergence of resistance but still our knowledge is inadequate. Proteins form a very complex network and drugs are countered by their modification/efflux or over expression/modification of targets. As proteins manifest most of the biological processes, these are attractive targets for developing drugs, immunodiagnostics or therapeutics. The aim of present study was to analyze and compare the protein profile of whole cell extracts from Mycobacterium tuberculosis clinical isolates susceptible and resistant to SM.ResultsTwo-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was employed for analyzing the protein profiles. Homology and in silico characterization for identified proteins was assessed using BLAST, InterProScan and KEGG database searches. Computational studies on the possible interactions between SM and identified proteins were carried out by a battery of online servers and softwares, namely, CLUSTALW (KEGG), I-TASSER, VMD, PatchDock and FireDock. On comparing 2DE patterns, nine proteins were found consistently overexpressed in SM resistant isolates and were identified as Rv0350, Rv0440, Rv1240, Rv3075c, Rv2971, Rv3028c, Rv2145c, Rv2031c and Rv0569. In silico docking analysis showed significant interactions of SM with essential (Rv0350, Rv0440 and Rv2971) and non essential (Rv1240, Rv3075c and Rv2031c) genes.ConclusionsThe computational results suggest high protein binding affinity of SM and suggested many possible interactions between identified proteins and the drug. Bioinformatic analysis proves attributive for analysis of diversity of proteins identified by whole proteome analysis. In-depth study of the these proteins will give an insight into probable sites of drug action other than established primary sites and hence may help in search of novel chemotherapeutic agents at these new sites as inhibitors.

Proteomic analysis of sensitive and multi drug resistant Mycobacterium tuberculosis strains

“…In another study, four proteins Rv0635, Rv1827, Rv0036c, and Rv2032 were identified as intracellular MDR-TB and two proteins of Rv2896c and Rv2558c were belonged to sensitive isolates (Neelja et al, 2012). One other study found nine proteins as overexpressed in SM-resistant isolates consisting of Rv0350, Rv0440, Rv1240, Rv3057c, Rv3028c, Rv2145c, Rv2031c, and Rv0569 (Sharma et al, 2010). These results are approximately similar to our findings.…”

“…Automated two-dimensional electrophoresis coupled with MALDI-TOF-TOF (matrix-assistedlaser desorption/ionization) has proven to be very efficient in the analysis of complex protein/peptide mixtures. In contrast to conventional protein sequencing procedures; mass spectrometry allows the rapid and automated analysis of many proteins in a short time frame (Shevchenko et al, 1996(Shevchenko et al, , 2006.…”

Synergistic antibacterial activity of the combination of the alkaloid sanguinarine with EDTA and the antibiotic streptomycin against multidrug resistant bacteria

“…The third drug in our study is the aminoglycoside antibiotic streptomycin, which targets protein synthesis by direct interaction with the small ribosomal subunit [11]. Streptomycin was the first drug used in the treatment of Mycobacterium tuberculosis and is preferred for treatment of relapses [12]…”

Resistome analysis of Mycobacterium tuberculosis: Identification of aminoglycoside 2’-N-acetyltransferase (AAC) as co-target for drug desigining

“…Coexpression and Occurrence analysis for this protein was obtained from STRING database. Blastp analysis [27] against a human protein database was done to validate that this particular protein not share any homology with human proteins.…”