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Comparative proteome analysis of culture supernatant proteins of Mycobacterium tuberculosis H37Rv and H37Ra.
- Source :
-
Microbes and infection [Microbes Infect] 2003 Aug; Vol. 5 (10), pp. 851-6. - Publication Year :
- 2003
-
Abstract
- To examine the virulence factors of Mycobacterium tuberculosis H37Rv, the proteome was used to characterize the differences in protein expression between virulent M. tuberculosis H37Rv and attenuated M. tuberculosis H37Ra. Two-dimensional gel electrophoresis was performed to separate culture supernatant proteins extracted from M. tuberculosis H37Rv and M. tuberculosis H37Ra. The protein spots of interest were identified by mass spectrometry, and then the genes encoding the identified proteins were cloned and sequenced. Comparison of silver-stained gels showed that three well-resolved protein spots were present in M. tuberculosis H37Rv but absent from M. tuberculosis H37Ra. Protein spot no. 1 was identified as Rv2346c. Protein spot no. 2 was identified as Rv2347c, Rv1197, Rv1038c, and Rv3620c, which shared significant homology and had the same peptide fingerprinting using tryptic digestion. No M. tuberculosis protein matched protein spot no. 3. Rv2346c, Rv2347c, Rv1038c, and Rv3620c of M. tuberculosis H37Rv were located on the M. tuberculosis H37Ra chromosome, and multiple mutations were observed in the corresponding areas of M. tuberculosis H37Ra. Codon 59 (CAG, Gln) of Rv2347c and Rv3620c was replaced by termination codon (TAG) in M. tuberculosis H37Ra, which probably terminated the polypeptide elongation. These results demonstrate the importance of studying the gene products of M. tuberculosis and show that subtle differences in isogenic mutant strains might play an important role in identifying the attenuating mutations.
- Subjects :
- Electrophoresis, Gel, Two-Dimensional
Hydrolysis
Mass Spectrometry
Mutation
Mycobacterium tuberculosis genetics
Mycobacterium tuberculosis pathogenicity
Peptide Mapping
Polymerase Chain Reaction
Protein Transport
Sequence Analysis, DNA
Silver Staining
Trypsin
Bacterial Proteins analysis
Mycobacterium tuberculosis metabolism
Proteome analysis
Virulence Factors analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1286-4579
- Volume :
- 5
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Microbes and infection
- Publication Type :
- Academic Journal
- Accession number :
- 12919853
- Full Text :
- https://doi.org/10.1016/s1286-4579(03)00179-5