Quantitative proteomics of microbes: Principles and applications to virulence

The rapidly increasing ability to sequence complete genomes of different microbial species and strains provides us with information regarding their genetic variability. Genetic variability is a mechanism for human pathogens to adapt to and avoid the immune system and to also develop resistance to antibiotics. However, the assessment of the contributions of individual genetic differences to resistance or other phenotypes is not a priori apparent from the genomic variability. Quantitative proteomics can provide accurate molecular phenotypes of microbes that are difficult to determine using alternative technologies. Over the recent few years we and others have developed a range of proteomic technologies for the quantitative analysis of microbial proteomes. Here, we describe the most commonly used techniques and discuss their strengths and weaknesses and illustrate their respective performance for the identification of virulence factors in Streptococcus pyogenes.