Progress with gene‐product mapping of the Mollicutes: Mycoplasma genitalium

Abstract

A protein map of the smallest known self-replicating organism, Mycoplasma genitalium (Class: Mollicutes), revealed a high proportion of acidic proteins. Amino acid composition was used to putatively identify, or provide unique parameters, for 50 gene products separated by two-dimensional gel electrophoresis. A further 19 proteins were subjected to peptide-mass fingerprinting using matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry and 4 were subjected to N-terminal Edman degradation. The majority of M. genitalium proteins remain uncharacterised. However, the combined approach of amino acid analysis and peptide-mass fingerprinting allowed gene products to be linked to homologous genes in a variety of organisms. This has allowed proteins to be identified prior to detection of their respective genes via the M. genitalium sequencing initiative. The principle of 'hierarchical' analysis for the mass screening of proteins and the analysis of microbial genomes via their protein complement or 'proteome' is detailed. Here, characterisation of gene products depends upon the quickest and most economical technologies being employed initially, so as to determine if a large number of proteins are already present in both homologous and heterologous species databases. Initial screening, which lends itself to automation and robotics, can then be followed by more time and cost intensive procedures, when necessary.

Guidelines for the next 10 years of proteomics

“…Proteomics by name is now over 10 years old [1,2] and many of the key technological innovations that made proteomics possible are of a similar or greater age. MS-based protein identification using PMF or LC-MS/MS [3][4][5] is now widely adopted, and the transition from analysing one protein at a time to analysing highly complex mixtures has been made [6,7].…”

Advances in proteomics for production strain analysis

“…F -scale study of proteins within a cell, tissue or organism [3]. It is a rapidly evolving field focused on identification and characterisation of these proteins and their proteoforms…”

Contribution of proteomics of Leishmania spp. to the understanding of differentiation, drug resistance mechanisms, vaccine and drug development

“…The first proteome maps of Leishmania were published in the early 1980s [9][10][11] long before the term proteomics was coined 12 . Handman et al as well as Saravia and colleagues were investigating species differences and virulence factors [9][10][11] .…”