Abstract
Mycoplasma fermentans is a potent human pathogen which has been implicated in several diseases. Notably, its lipid-associated membrane proteins (LAMPs) play a role in immunomodulation and development of infection-associated inflammatory diseases. However, the systematic protein identification of pathogenic M. fermentans has not been reported. From our recent sequencing results of M. fermentans M64 isolated from human respiratory tract, its genome is around 1.1 Mb and encodes 1050 predicted protein-coding genes. In the present study, soluble proteome of M. fermentans was resolved and analyzed using two-dimensional gel electrophoresis. In addition, Triton X-114 extraction was carried out to enrich amphiphilic proteins including putative lipoproteins and membrane proteins. Subsequent mass spectrometric analyses of these proteins had identified a total of 181 M. fermentans ORFs. Further bioinformatics analysis of these ORFs encoding proteins with known or so far unknown orthologues among bacteria revealed that a total of 131 proteins are homologous to known proteins, 11 proteins are conserved hypothetical proteins, and the remaining 39 proteins are likely M. fermentans-specific proteins. Moreover, Triton X-114-enriched fraction was shown to activate NF-kB activity of raw264.7 macrophage and a total of 21 lipoproteins with predicted signal peptide were identified therefrom. Together, our work provides the first proteome reference map of M. fermentans as well as several putative virulence-associated proteins as diagnostic markers or vaccine candidates for further functional study of this human pathogen.
Highlights
Mycoplasmas belong to the Mollicutes class of organisms and are often considered as wall-less bacteria with the smallest genomes
The bacteria can import nucleosides, amino acids and metabolites from the rich growth media, it is anticipated that only a minimum amount of proteins were expressed [38]
Our results showed that the proteins in Triton X-114 extraction showed a significant NF-kB activation activity in mouse macrophage cells compared to the aqueous fraction. (p = 0.001082)
Summary
Mycoplasmas belong to the Mollicutes class of organisms and are often considered as wall-less bacteria with the smallest genomes. Colonization of Mollicutes has been found in both plants and animals, where many animal mycoplasmas are extracellular parasites and plant mycoplasmas are localized solely in the phloem sieve tubes of affected plants and transmitted by insect vectors [1,2]. Scientists have isolated at least 16 species of Mycoplasma from humans and their major colonization sites include oropharynx, upper respiratory tract, and genitourinary tract [1]. Mycoplasma fermentans, a human cell-invasive mycoplasma, has been suspected to associate with several human diseases. Presence of M. fermentans among acquired immunodeficiency syndrome (AIDS) patients was reported in 1989
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