Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis

Itay Raphael, Swetha Mahesula, Karan Kalsaria, Venkat Kotagiri, Anjali B. Purkar, Manjushree Anjanappa, Darshit Shah, Vidya Pericherla, Yeshwant Lal Avinash Jadhav, Rekha Raghunathan, Michael Vaynberg, David Noriega, Nazul H. Grimaldo, Carola Wenk, Jonathan A.L. Gelfond, Thomas G. Forsthuber, William E. Haskins

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12 Scopus citations

Abstract

We hypothesized that quantitative MS/MS-based proteomics at multiple time points, incorporating rapid microwave and magnetic (M2) sample preparation, could enable relative protein expression to be correlated to disease progression in the experimental autoimmune encephalomyelitis (EAE) animal model of multiple sclerosis. To test our hypothesis, microwave-assisted reduction/alkylation/digestion of proteins from brain tissue lysates bound to C8 magnetic beads and microwave-assisted isobaric chemical labeling were performed of released peptides, in 90 s prior to unbiased proteomic analysis. Disease progression in EAE was assessed by scoring clinical EAE disease severity and confirmed by histopathologic evaluation for central nervous system inflammation. Decoding the expression of 283 top-ranked proteins (p <0.05) at each time point relative to their expression at the peak of disease, from a total of 1191 proteins observed in four technical replicates, revealed a strong statistical correlation to EAE disease score, particularly for the following four proteins that closely mirror disease progression: 14-3-3ε (p = 3.4E-6); GPI (p = 2.1E-5); PLP1 (p = 8.0E-4); PRX1 (p = 1.7E-4). These results were confirmed by Western blotting, signaling pathway analysis, and hierarchical clustering of EAE risk groups. While validation in a larger cohort is underway, we conclude that M2 proteomics is a rapid method to quantify putative prognostic/predictive protein biomarkers and therapeutic targets of disease progression in the EAE animal model of multiple sclerosis.

Original languageEnglish (US)
Pages (from-to)3810-3819
Number of pages10
JournalELECTROPHORESIS
Volume33
Issue number24
DOIs
StatePublished - Dec 2012

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Keywords

  • Isobaric chemical labeling
  • Microwave proteomics
  • Multiple sclerosis
  • Sample preparation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this

Raphael, I., Mahesula, S., Kalsaria, K., Kotagiri, V., Purkar, A. B., Anjanappa, M., Shah, D., Pericherla, V., Jadhav, Y. L. A., Raghunathan, R., Vaynberg, M., Noriega, D., Grimaldo, N. H., Wenk, C., Gelfond, J. A. L., Forsthuber, T. G., & Haskins, W. E. (2012). Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis. ELECTROPHORESIS, 33(24), 3810-3819. https://doi.org/10.1002/elps.201200200