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A two\u2010dimensional electrophoresis gel reference map of <jats:italic>S. melliferum<\/jats:italic> was produced by Phoretix 2\u2010D gel software analysis of eight high quality gels. The reference map showed 456 silver\u2010stained and replicated protein spots. 156 proteins (34% of visible protein spots) from <jats:italic>S. melliferum<\/jats:italic> were further characterised by one, or a combination, of the following: amino acid analysis, peptide\u2010mass fingerprinting <jats:italic>via<\/jats:italic> matrix assisted laser desorption ionisation\u2010time of light (MALDI\u2010TOF) mass spectrometry, and <jats:italic>N<\/jats:italic>\u2010terminal 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\u2010scale analysis of a proteome based upon the attribution of predominantly \u2018unique numerical parameters\u2019 for protein characterisation across species boundaries, as opposed to a sequence\u2010based 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. Geneproducts from major families such as glycolysis, translation, transcription, cellular processes, energy metabolism and protein synthesis were identified. Several gene\u2010products characterised in <jats:italic>S. melliferum<\/jats:italic> were not previously found in studies of the entire <jats:italic>Mycoplasma genitalium<\/jats:italic> and <jats:italic>Mycoplasma pneumoniae<\/jats:italic> (both closely related Mollicutes) genomes. The presence of such geneproducts in <jats:italic>S. melliferum<\/jats:italic> is discussed in terms of genome size as compared with the smallest known free\u2010living organisms. Finally, the levels of expression of <jats:italic>S. melliferum<\/jats:italic> gene\u2010products were determined with respect to total optical itensity associated with all visible proteins expressed in exponentially grown cells.<\/jats:p>","DOI":"10.1002\/elps.1150180809","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T12:14:38Z","timestamp":1117282478000},"page":"1335-1346","source":"Crossref","is-referenced-by-count":49,"title":["Proteome analysis of <i>Spiroplasma melliferum<\/i> (A56) and protein characterisation across species boundaries"],"prefix":"10.1002","volume":"18","author":[{"given":"Stuart J.","family":"Cordwell","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David 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