Author:
Sharma Prashant,Kumar Bhavnesh,Gupta Yash,Singhal Neelja,Katoch Vishwa Mohan,Venkatesan Krishnamurthy,Bisht Deepa
Abstract
Abstract
Background
Streptomycin (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.
Results
Two-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.
Conclusions
The 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.
Publisher
Springer Science and Business Media LLC
Subject
Molecular Biology,Biochemistry
Reference41 articles.
1. World Health Organization: Global tuberculosis control: epidemiology, strategy, financing. WHO Report 2009. [http://www.who.int/tb/publications/global_report/2009/pdf/full_report.pdf]
2. Franco H, Wowk PF, Silva CL, Trombone APF, Coelho-Castelo AAM, Oliver C, Jamur MC, Moretto EL, Bonato VLD: A DNA vaccine against tuberculosis based on the 65 kDa heat-shock protein differentially activates human macrophages and dendritic cells. Genet Vaccines Ther 2008, 6: 3. 10.1186/1479-0556-6-3
3. Zhang Y, Telenti A: Genetics of drug resistance in Mycobacterium tuberculosis . In Molecular Genetics of Mycobacteria. Edited by: Hatfull GF, Jacobs WR. Washington DC: ASM Press; 2000:235–254.
4. Honore N, Cole ST: Streptomycin resistance in mycobacteria. Antimicrob Agents Chemother 1994,38(2):238–242.
5. Ulger M, Aslan G, Emekdas G, Tezcan S, Serin MS: Investigation of rpsL and rrs gene region mutations in streptomycin resistant Mycobacterium tuberculosis complex isolates. Mikrobiyol Bul 2009,43(1):115–120.
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