Proteomic analysis and immunogenicity of secreted proteins from Rhodococcus equi ATCC 33701.
Abstract: Rhodococcus equi is one of the most important causes of mortality in foals between 1 and 6 months of age. Although rare, infection also occurs in a variety of other mammals including humans, often following immunosuppression of various causes. Secreted proteins are known to mediate important pathogen-host interactions and consequently are favored candidates for vaccine development as they are the most easily accessible microbial antigens to the immune system. Here, we describe the results of a proteomic analysis based on SDS-PAGE, immunoblot and mass spectrometry, which was carried out aiming the identification of secreted proteins that are differently expressed at 30 degrees C versus 37 degrees C and at mid-exponential versus early-stationary growth phase and antigenic proteins from R. equi ATCC 33701. A total of 48 proteins was identified regardless of growth conditions. The cholesterol oxidase ChoE appears to be the major secretory protein. Moreover, four proteins revealed high homologies with the mycolyl transferases of the Ag85 complex from Mycobacterium tuberculosis. The sequence analysis predicted that 24 proteins are transported by a signal peptide-dependent pathway. Moreover, five antigenic proteins of R. equi were identified by immunoblot, including a novel strongly immunoreactive protein of unknown function. In conclusion, the elucidation of the secretome of R. equi identified several proteins with different biological functions and a new candidate for developing vaccines against R. equi infection in horse.
Publication Date: 2008-10-15 PubMed ID: 19019579DOI: 10.1016/j.vetmic.2008.09.086Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study examines the proteins secreted by Rhodococcus equi, a harmful bacteria to young foals and other mammals, to determine potential candidates for vaccine development. The research identified several unique proteins and identified a previously unknown, strongly immunoreactive protein which could serve as a candidate for new vaccine development.
Research Scope and Methods
- The study identifies secreted proteins of Rhodococcus equi, a pathogen responsible for the death of foals between 1 and 6 months old and an infectious agent in other mammals, including humans, post-immunosuppression.
- The researchers engaged in a proteomic analysis involving Sodium Dodecyl Sulfate – Polyacrylamide Gel Electrophoresis (SDS-PAGE), immunoblot and mass spectrometry to identify these secreted proteins and how they differ under different conditions of temperature and growth phase.
Findings
- The research identified a total of 48 proteins that R. equi secreted, irrespective of the growth conditions. Among these, the cholesterol oxidase ChoE seemed to be the predominant secretory protein.
- Four of these proteins were found to be greatly similar to the mycolyl transferases of the Ag85 complex from Mycobacterium tuberculosis. Mycolyl transferases are involved in the biosynthesis of mycolic acids, which are vital components of the Mycobacterium cell wall, so these proteins caught the researchers’ attention.
- Based on sequence analysis, it was predicted that 24 proteins are transported by a pathway that is dependent on signal peptides.
- Analysis of antigens by immunoblot revealed five immunogenic proteins in R. equi. One protein, previously not known, showed strong immunoreactivity, making it of special interest.
Significance and Implication
- The research enhances the understanding of the secretome (the group of secreted proteins) of R. equi, revealing several proteins that can be associated with different biological functions.
- Prominently, the elucidation of the secretome brings forth a new candidate protein, displaying strong immunoreactivity, that might be utilized in developing vaccines against the infection caused by R. equi in horses.
- Recognizing these proteins is crucial because they can serve as the most accessible targets for the immune system to develop immunity, and hence can be instrumental in vaccine development.
Cite This Article
APA
Barbey C, Budin-Verneuil A, Cauchard S, Hartke A, Laugier C, Pichereau V, Petry S.
(2008).
Proteomic analysis and immunogenicity of secreted proteins from Rhodococcus equi ATCC 33701.
Vet Microbiol, 135(3-4), 334-345.
https://doi.org/10.1016/j.vetmic.2008.09.086 Publication
Researcher Affiliations
- AFSSA, Laboratoire d'Etudes et de Recherches en Pathologie Equine, IPC, Goustranville, 14430 Dozulé, France.
MeSH Terms
- Actinomycetales Infections / veterinary
- Amino Acid Sequence
- Animals
- Antigens, Bacterial / genetics
- Antigens, Bacterial / immunology
- Antigens, Bacterial / isolation & purification
- Bacterial Proteins / genetics
- Bacterial Proteins / immunology
- Bacterial Proteins / isolation & purification
- DNA Primers
- Electrophoresis, Polyacrylamide Gel
- Enzymes / chemistry
- Horse Diseases / microbiology
- Horses
- Immunoblotting
- Mass Spectrometry
- Mycobacterium tuberculosis / enzymology
- Mycobacterium tuberculosis / genetics
- Proteome
- Rhodococcus equi / genetics
- Rhodococcus equi / growth & development
- Rhodococcus equi / immunology
- Sequence Homology, Amino Acid
- Temperature
- Trypsin
Citations
This article has been cited 7 times.- Madec S, Pichereau V, Jacq A, Paillard M, Boisset C, Guérard F, Paillard C, Nicolas JL. Characterization of the secretomes of two vibrios pathogenic to mollusks. PLoS One 2014;9(11):e113097.
- Choi CW, An HY, Lee YJ, Lee YG, Yun SH, Park EC, Hong Y, Kim GH, Park JE, Baek SJ, Kim HS, Kim SI. Characterization of Streptococcus pneumoniae N-acetylglucosamine-6-phosphate deacetylase as a novel diagnostic marker. J Microbiol 2013 Oct;51(5):659-64.
- Ciprandi A, da Silva WM, Santos AV, de Castro Pimenta AM, Carepo MS, Schneider MP, Azevedo V, Silva A. Chromobacterium violaceum: important insights for virulence and biotechnological potential by exoproteomic studies. Curr Microbiol 2013 Jul;67(1):100-6.
- López E, López I, Ferreira A, Sequí J. Clinical and technical phosphoproteomic research. Proteome Sci 2011 Jun 2;9:27.
- Pacheco LG, Slade SE, Seyffert N, Santos AR, Castro TL, Silva WM, Santos AV, Santos SG, Farias LM, Carvalho MA, Pimenta AM, Meyer R, Silva A, Scrivens JH, Oliveira SC, Miyoshi A, Dowson CG, Azevedo V. A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis. BMC Microbiol 2011 Jan 17;11(1):12.
- Ranganathan S, Garg G. Secretome: clues into pathogen infection and clinical applications. Genome Med 2009 Nov 30;1(11):113.
- Paria P, Chakraborty HJ, Pakhira A, Devi MS, Das Mohapatra PK, Behera BK. Identification of virulence-associated factors in Vibrio parahaemolyticus with special reference to moonlighting protein: a secretomics study. Int Microbiol 2024 Jun;27(3):765-779.
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