Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: May 20, 2006
Publication Date: May 20, 2006
Citation: Radosevich, T.J., Stabel, J.R., Bannantine, J.P., Lippolis, J.D., Reinhardt, T.A. 2006. Proteome and Differential Expression Analysis of Membrane and Cytosolic Protein from Mycobacterium avium subs paratuberculosis Strains k10 and 187 [abstract]. American Society for Microbiology. p. 112. Technical Abstract: Paratuberculosis (Johne's disease) is a significant economic problem in cattle and sheep worldwide. The causative agent of the chronic enteric disease is Mycobacterium avium subsp. paratuberculosis (MAP). Little is known of the protein expression profile of MAP and how this contributes to pathogenesis. In the present study, proteins from both outer membranes and cytosol were prepared from two strains of MAP, laboratory strain K10 and a low passage isolate obtained from a clinical cow, strain 187. One-dimensional SDS Page and 2-dimensional blue native electrophoresis of membranes proteins from MAP strains K10 and 187 showed marked differences in protein expression between the 2 strains. Following this we measured relative proteins expression from both MAP strains using amine-reactive isobaric tagging reagents (iTRAQ) and tandem mass spectroscopy. Protein identifications were obtained for greater than 500 membrane and cytosolic proteins from the MAP proteome with relative protein expression data for these identified proteins. The data showed a number of significant differences in protein expression between the laboratory strain and the clinical isolate. Examples of proteins down regulated in strain 187 are RpmD, MAP0630c and MAP2433. In contrast, proteins such as Tuf and DesA1 were up regulated in strain 187 as compared to strain K10. These data may provide insights into the proteins whose expression is important in natural infection but are modified once MAP is subjected to a laboratory environment. If so, they will provide tools for developing a better understanding of MAP infection in the host, and offer potential as diagnostic reagents and vaccine candidates.