The liposoluble proteome of Mycoplasma agalactiae: an insight into the minimal protein complement of a bacterial membrane
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Data
2010-08-25
Autori
Cacciotto, Carla
Addis, Maria Filippa
Pagnozzi, Daniela
Chessa, Bernardo
Coradduzza, Elisabetta
Carcangiu, Laura
Uzzau, Sergio
Alberti, Alberto
Pittau, Marco
Titolo del periodico
ISSN
Titolo del volume
Editore
BioMed Central
Abstract
Background: Mycoplasmas are the simplest bacteria capable of autonomous replication. Their evolution
proceeded from gram-positive bacteria, with the loss of many biosynthetic pathways and of the cell wall. In this
work, the liposoluble protein complement of Mycoplasma agalactiae, a minimal bacterial pathogen causing
mastitis, polyarthritis, keratoconjunctivitis, and abortion in small ruminants, was subjected to systematic
characterization in order to gain insights into its membrane proteome composition.
Results: The selective enrichment for M. agalactiae PG2T liposoluble proteins was accomplished by means of Triton
X-114 fractionation. Liposoluble proteins were subjected to 2-D PAGE-MS, leading to the identification of 40 unique
proteins and to the generation of a reference 2D map of the M. agalactiae liposoluble proteome. Liposoluble
proteins from the type strain PG2 and two field isolates were then compared by means of 2D DIGE, revealing
reproducible differences in protein expression among isolates. An in-depth analysis was then performed by GeLCMS/
MS in order to achieve a higher coverage of the liposoluble proteome. Using this approach, a total of 194
unique proteins were identified, corresponding to 26% of all M. agalactiae PG2T genes. A gene ontology analysis
and classification for localization and function was also carried out on all protein identifications. Interestingly, the
11.5% of expressed membrane proteins derived from putative horizontal gene transfer events.
Conclusions: This study led to the in-depth systematic characterization of the M. agalactiae liposoluble protein
component, providing useful insights into its membrane organization.
Descrizione
Keywords
Mycoplasma agalactiae , liposoluble proteins , PG2T genes