Proteome analysis ofSpiroplasma melliferum (A56) and protein characterisation across species boundaries
| Autor: | David J. Basseal, Ian Humphery-Smith, Stuart J. Cordwell |
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| Rok vydání: | 1997 |
| Předmět: |
Sequence analysis
Spiroplasma Molecular Sequence Data Clinical Biochemistry Gene Expression Computational biology Peptide Mapping Biochemistry Genome Analytical Chemistry Protein sequencing Bacterial Proteins Species Specificity Electrophoresis Gel Two-Dimensional Amino Acid Sequence Peptide sequence Genome size Genetics biology biology.organism_classification Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Proteome Mollicutes Sequence Analysis Genome Bacterial |
| Zdroj: | Electrophoresis. 18:1335-1346 |
| ISSN: | 1522-2683 0173-0835 |
| DOI: | 10.1002/elps.1150180809 |
| Popis: | Spiroplasma melliferum (Class: Mollicutes) is a wall-less, helical bacterium with a genome of approximately 1460 kbp encoding 800-1000 gene-products. A two-dimensional electrophoresis gel reference map of S. melliferum was produced by Phoretix 2-D gel software analysis of eight high quality gels. The reference map showed 456 silver-stained and replicated protein spots. 156 proteins (34% of visible protein spots) from S. melliferum were further characterised by one, or a combination, of the following: amino acid analysis, peptide-mass fingerprinting via matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry, and N-terminal protein microsequencing. Proteins with close relationship to those previously determined from other species were identified across species barriers. Thus, this study represents the first larger-scale analysis of a proteome based upon the attribution of predominantly 'unique numerical parameters' for protein characterisation across species boundaries, as opposed to a sequence-based approach. This approach allowed all database entries to be screened for homology, as is currently the case for studies based on nucleic acid or protein sequence information. Several proteins studied from this organism were identified as hypothetical, or having no close homolog already present in the databases. Gene-products from major families such as glycolysis, translation, transcription, cellular processes, energy metabolism and protein synthesis were identified. Several gene-products characterised in S. melliferum were not previously found in studies of the entire Mycoplasma genitalium and Mycoplasma pneumoniae (both closely related Mollicutes) genomes. The presence of such gene-products in S. melliferum is discussed in terms of genome size as compared with the smallest known free-living organisms. Finally, the levels of expression of S. melliferum gene-products were determined with respect to total optical intensity associated with all visible proteins expressed in exponentially grown cells. |
| Databáze: | OpenAIRE |
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