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Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis
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 o...
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| Published in: | Electrophoresis 2012-12, Vol.33 (24), p.3810-3819 |
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| creator | Raphael, Itay Mahesula, Swetha Kalsaria, Karan Kotagiri, Venkat Purkar, Anjali B. Anjanappa, Manjushree Shah, Darshit Pericherla, Vidya Jadhav, Yeshwant Lal Avinash Raghunathan, Rekha Vaynberg, Michael Noriega, David Grimaldo, Nazul H. Wenk, Carola Gelfond, Jonathan A.L. Forsthuber, Thomas G. Haskins, William E. |
| description | 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 |
| doi_str_mv | 10.1002/elps.201200200 |
| format | article |
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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.</description><identifier>ISSN: 0173-0835</identifier><identifier>EISSN: 1522-2683</identifier><identifier>DOI: 10.1002/elps.201200200</identifier><identifier>PMID: 23161666</identifier><language>eng</language><publisher>Germany: Blackwell Publishing Ltd</publisher><subject>Animals ; Beads ; Blotting, Western ; Brain - metabolism ; Brain - physiopathology ; Brain Chemistry ; Central nervous system ; Cluster Analysis ; Disease Models, Animal ; Encephalomyelitis, Autoimmune, Experimental - metabolism ; Female ; Isobaric chemical labeling ; Magnetics ; Mice ; Mice, Inbred C57BL ; Microwave proteomics ; Microwaves ; Multiple sclerosis ; Multiple Sclerosis - metabolism ; Progressions ; Proteins ; Proteome - analysis ; Proteome - metabolism ; Proteomics ; Proteomics - methods ; Sample preparation ; Tandem Mass Spectrometry - methods</subject><ispartof>Electrophoresis, 2012-12, Vol.33 (24), p.3810-3819</ispartof><rights>2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><rights>2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5062-aa7d4ebff3a95ad76f330b2b4bb0df2eff6c47819c2287444b06c7e4b69793873</citedby><cites>FETCH-LOGICAL-c5062-aa7d4ebff3a95ad76f330b2b4bb0df2eff6c47819c2287444b06c7e4b69793873</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23161666$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Raphael, Itay</creatorcontrib><creatorcontrib>Mahesula, Swetha</creatorcontrib><creatorcontrib>Kalsaria, Karan</creatorcontrib><creatorcontrib>Kotagiri, Venkat</creatorcontrib><creatorcontrib>Purkar, Anjali B.</creatorcontrib><creatorcontrib>Anjanappa, Manjushree</creatorcontrib><creatorcontrib>Shah, Darshit</creatorcontrib><creatorcontrib>Pericherla, Vidya</creatorcontrib><creatorcontrib>Jadhav, Yeshwant Lal Avinash</creatorcontrib><creatorcontrib>Raghunathan, Rekha</creatorcontrib><creatorcontrib>Vaynberg, Michael</creatorcontrib><creatorcontrib>Noriega, David</creatorcontrib><creatorcontrib>Grimaldo, Nazul H.</creatorcontrib><creatorcontrib>Wenk, Carola</creatorcontrib><creatorcontrib>Gelfond, Jonathan A.L.</creatorcontrib><creatorcontrib>Forsthuber, Thomas G.</creatorcontrib><creatorcontrib>Haskins, William E.</creatorcontrib><title>Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis</title><title>Electrophoresis</title><addtitle>ELECTROPHORESIS</addtitle><description>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.</description><subject>Animals</subject><subject>Beads</subject><subject>Blotting, Western</subject><subject>Brain - metabolism</subject><subject>Brain - physiopathology</subject><subject>Brain Chemistry</subject><subject>Central nervous system</subject><subject>Cluster Analysis</subject><subject>Disease Models, Animal</subject><subject>Encephalomyelitis, Autoimmune, Experimental - metabolism</subject><subject>Female</subject><subject>Isobaric chemical labeling</subject><subject>Magnetics</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Microwave proteomics</subject><subject>Microwaves</subject><subject>Multiple sclerosis</subject><subject>Multiple Sclerosis - metabolism</subject><subject>Progressions</subject><subject>Proteins</subject><subject>Proteome - analysis</subject><subject>Proteome - metabolism</subject><subject>Proteomics</subject><subject>Proteomics - methods</subject><subject>Sample preparation</subject><subject>Tandem Mass Spectrometry - methods</subject><issn>0173-0835</issn><issn>1522-2683</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNqNkc1v1DAQxS0EokvhyhHlWA5Z_BU7viDRqrSgLRTxdbQcZ9I12HGIk7bLX49XW1ZwgpM1nt97mpmH0FOClwRj-gL8kJYUE5oLjO-hBakoLamo2X20wESyEtesOkCPUvqGMeaK84fogDIiiBBigX5eODvGG3MNhenbIpirHiZni6ML-rwYxjhBDM6mInbFtIYCbgcYXYB-Mr4w8xRdCHOf_3sLw9r4GDbg3eRSdnMhMyG24LfqMPvJDR6KZD2MMbn0GD3ojE_w5O49RJ9fn346OS9X78_enLxalbbCgpbGyJZD03XMqMq0UnSM4YY2vGlw21HoOmG5rImylNaSc95gYSXwRiipWC3ZIXq58x3mJkBr8_Cj8XrIe5hxo6Nx-u9O79b6Kl5rJjGTAmeDozuDMf6YIU06uGTBe9NDnJMmnCkplOL_gVLJqKRcVBld7tB8_5RG6PYTEay32epttnqfbRY8-3OPPf47zAzwHXDjPGz-YadPV5cf87VolpU7mUsT3O5lZvyuhWSy0l_fnelzeqy-kMu3-gP7BV-hwk4</recordid><startdate>201212</startdate><enddate>201212</enddate><creator>Raphael, Itay</creator><creator>Mahesula, Swetha</creator><creator>Kalsaria, Karan</creator><creator>Kotagiri, Venkat</creator><creator>Purkar, Anjali B.</creator><creator>Anjanappa, Manjushree</creator><creator>Shah, Darshit</creator><creator>Pericherla, Vidya</creator><creator>Jadhav, Yeshwant Lal Avinash</creator><creator>Raghunathan, Rekha</creator><creator>Vaynberg, Michael</creator><creator>Noriega, David</creator><creator>Grimaldo, Nazul H.</creator><creator>Wenk, Carola</creator><creator>Gelfond, Jonathan A.L.</creator><creator>Forsthuber, Thomas G.</creator><creator>Haskins, William E.</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7U5</scope><scope>8FD</scope><scope>L7M</scope><scope>5PM</scope></search><sort><creationdate>201212</creationdate><title>Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis</title><author>Raphael, Itay ; 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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.</abstract><cop>Germany</cop><pub>Blackwell Publishing Ltd</pub><pmid>23161666</pmid><doi>10.1002/elps.201200200</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Electrophoresis, 2012-12, Vol.33 (24), p.3810-3819 |
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| subjects | Animals Beads Blotting, Western Brain - metabolism Brain - physiopathology Brain Chemistry Central nervous system Cluster Analysis Disease Models, Animal Encephalomyelitis, Autoimmune, Experimental - metabolism Female Isobaric chemical labeling Magnetics Mice Mice, Inbred C57BL Microwave proteomics Microwaves Multiple sclerosis Multiple Sclerosis - metabolism Progressions Proteins Proteome - analysis Proteome - metabolism Proteomics Proteomics - methods Sample preparation Tandem Mass Spectrometry - methods |
| title | Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis |
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