The Low Molecular Weight Proteome of Halobacterium salinarum

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• 作者：Christian Klein ; Michalis Aivaliotis ; Jesper V. Olsen ; Michaela Falb ; H&uuml ; seyin Besir ; Beatrix Scheffer ; Birgit Bisle ; Andreas Tebbe ; Kosta Konstantinidis ; Frank Siedler ; Friedhelm Pfeiffer ; Matthi
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：April 2007
• 年：2007
• 卷：6
• 期：4
• 页码：1510 - 1518
• 全文大小：602K
• 年卷期：v.6,no.4(April 2007)
• ISSN：1535-3907

文摘

Systematic investigation of low molecular weight proteins (LMW, below 20 kDa) in the archaeonHalobacterium salinarum resulted in a 6-fold enhancement of the identification rate, reaching 35% ofthe theoretical proteome in that size range. This was achieved by optimization of common protocolsfor protein analysis with general applicability. LMW proteins were rapidly and effectively enriched byfilter membrane centrifugation followed by tricine SDS-PAGE. Without staining and with significantlyshortened digestion protocols, LMW proteins were identified using an FT-ICR mass spectrometer whichallows reliable protein identification by MS³ of a single peptide. In addition to a series of technicalchallenges, small proteins may show low gene expression levels as suggested by their low averagecodon adaptation index. Twenty functionally uncharacterized proteins contain a characteristic DNA/RNA binding zinc finger motif which underlines the biological relevance of the small proteome andthe necessity of their analysis for systems biology.