Abstract
It is now more than 10 years since the publication of the first microbial genome sequence and science is now moving towards a post genomic era with transcriptomics and proteomics offering insights into cellular processes and function. The ability to assess the entire protein network of a cell at a given spatial or temporal point will have a profound effect upon microbial science as the function of proteins is inextricably linked to phenotype. Whilst such a situation is still beyond current technologies rapid advances in mass spectrometry, bioinformatics and protein separation technologies have produced a step change in our current proteomic capabilities. Subsequently a small, but steadily growing, number of groups are taking advantage of this cutting edge technology to discover more about the physiology and metabolism of microorganisms. From this research it will be possible to move towards a systems biology understanding of a microorganism. Where upon researchers can build a comprehensive cellular map for each microorganism that links an accurately annotated genome sequence to gene expression data, at a transcriptomic and proteomic level.In order for microbiologists to embrace the potential that proteomics offers, an understanding of a variety of analytical tools is required. The aim of this review is to provide a basic overview of mass spectrometry (MS) and its application to protein identification. In addition we will describe how the protein complexity of microbial samples can be reduced by gel-based and gel-free methodologies prior to analysis by MS. Finally in order to illustrate the power of microbial proteomics a case study of its current application within the Bacilliaceae is given together with a description of the emerging discipline of metaproteomics.
Highlights
Mass spectrometry has its origins in the studies performed by J
Microorganisms are ideal candidates for systems biology research and the field of systems microbiology is expected to result in the development of tools and techniques of general applicability across the life sciences [106]
Proteomics has a key role to play in attempts to construct comprehensive cellular maps of biochemical processes occurring within specific microorganisms at given spatial and temporal points
Summary
Mass spectrometry has its origins in the studies performed by J. The chip has a number of sample wells each containing a different chemistry, specific classes of molecules may be captured from cell lysates and selectively analyzed Using this process several toxigenic and nontoxigenic strains of Bacteroides fragilis were analyzed revealing potential biomarkers specific to the toxigenic strains in the mass range 3.5–18.5 kDa. Expressional Proteomics Whilst techniques described far provide the microbiologist with an invaluable snapshot of the processes occurring within a biological system, assessing the quantitative change in protein expression patterns remains the focus for those interested in the fundamental analysis of microbial systems. Digest proteins from up to four states A B C D
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