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Identifying Targets for Therapeutic Interventions using Proteomic Technology


EMSL Project ID
10904a

Abstract

This proposal will utilize new comprehensive and quantitative proteomic technologies and related capabilities developed at PNNL to identify targets for therapeutic intervention for Salmonella typhimurium, Salmonella typhi, the orthopox virus particles for vaccina, and monkey pox.

Preliminary target identification from the bacteria will be the result of comparison of protein expression profiles of wild type and regulatory mutants in pure culture and in macrophage, dendritic epithelial and fibroblast host cell lines. The comparison of these profiles using bioinformatics approaches will then serve to identify protein targets important for replication and pathogenesis. Additional validation of not only the target proteins but also the bioinformatics approach will be accomplished though the screening of mutations made in target proteins.

Preliminary target identification in the virus particles will be accomplished by comparison of both variola virus (VV) and monkey pox virus (MPV) in Hela and THP-differentiated cell lines where the difference in protein expression patterns will elucidate the proteins important in pathogenesis. Once the bioinformatics approach has been demonstrated and validated with Salmonella, the approach will be extended to proteomic studies of orthopox virus particles for the early validation of the protein targets.

Project Details

Start Date
2007-08-13
End Date
2010-08-15
Status
Closed

Team

Principal Investigator

Joshua Adkins
Institution
Pacific Northwest National Laboratory

Team Members

Roslyn Brown
Institution
Washington State University Tri-Cities

Charles Ansong
Institution
National Institutes of Health

Joseph Brown
Institution
Pacific Northwest National Laboratory

Saiful Chowdhury
Institution
Pacific Northwest National Laboratory

Errol Robinson
Institution
Pacific Northwest National Laboratory

Heather Olson
Institution
Environmental Molecular Sciences Laboratory

Fred Heffron
Institution
Oregon Health & Science University

Karin Rodland
Institution
Pacific Northwest National Laboratory

Related Publications

Ansong, C.K., Norbeck, A.D., Yoon, H., Gustin, J.K., McDermott, J.E., Mottaz, H.M., Adkins, J.N., Heffron, F. & Smith, R.D. Proteomics Analysis of the Cau sative Agent of Typhoid Fever. J Proteome Res (2008).
Ansong, C., Purvine, S.O., Adkins, J.N., Lipton, M.S. & Smith, R.D. Proteogenomics: needs and roles to be filled by proteomics in genome annotation. Brief Funct Genomic Proteomic 7, 50-62 (2008).
Brown JN, R Estep, D Lopez-Ferrer, HM Brewer, TRW Clauss, NP Manes, M O'Connor, H Li, JN Adkins, S Wong, and RD Smith. 2010. "CHARACTERIZATION OF MACAQUE PULMONARY FLUID PROTEOME DURING MONKEYPOX INFECTION: DYNAMICS OF HOST RESPONSE." Molecular & Cellular Proteomics. MCP 9(12):2760-2771. doi:10.1074/mcp.M110.001875
Callister, S.J., McCue, L.A., Turse, J.E., Monroe, M.E., Auberry, K.J., Smith, R.D., Adkins, J.N. & Lipton, M.S. Comparative bacterial proteomics: analysis of the core genome concept. PLoS ONE 3, e1542 (2008).
Chowdhury SM, L Shi, H Yoon, C Ansong, LM Rommereim, AD Norbeck, KJ Auberry, RJ Moore, JN Adkins, F Heffron, and RD Smith. 2009. "A method for investigating protein-protein interactions related to Salmonella typhimurium pathogenesis ." Journal of Proteome Research 8(3):1504-1514
Du, X., Callister, S.J., Manes, N.P., Adkins, J.N., Alexandridis, R.A., Zeng, X., Roh, J.H., Smith, W.E., Donohue, T.J., Kaplan, S., Smith, R.D. & Lipton, M.S. A Computational Strategy to Analyze Label-Free Temporal Bottom-Up Proteomics Data. J Proteome Res (2008).
Manes, N.P., Estep, R.D., Mottaz, H.M., Moore, R.J., Clauss, T.R., Monroe, M.E., Du, X., Adkins, J.N., Wong, S.W. & Smith, R.D. Comparative Proteomics of Human Monkeypox and Vaccinia Intracellular Mature and Extracellular Enveloped Virions. J Proteome Res (2008).
Manes, N.P., Gustin, J.K., Rue, J., Mottaz, H.M., Purvine, S.O., Norbeck, A.D., Monroe, M.E., Zimmer, J.S., Metz, T.O., Adkins, J.N., Smith, R.D. & Heffron, F. Targeted Protein Degradation by Salmonella under Phagosome-mimicking Culture Conditions Investigated Using Comparative Peptidomics. Mol Cell Proteomics 6, 717-27 (2007).
Rodland KD, JN Adkins, CK Ansong, SM Chowdhury, NP Manes, L Shi, H Yoon, RD Smith, and F Heffron. 2008. "Use of high-throughput mass spectrometry to elucidate host pathogen interactions in Salmonella." Future Microbiology
Yoon H, C Ansong, JE McDermott, MA Gritsenko, RD Smith, F Heffron, and JN Adkins. 2011. "Systems analysis of multiple regulator perturbations allows discovery of virulence factors in Salmonella ." BMC Systems Biology 5:Article No. 100. doi:10.1186/1752-0509-5-100