| Uncovering the proteome response of the master circadian clock to light using an AutoProteome system. | |
MedLine Citation:
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PMID: 21859948 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In mammals, the suprachiasmatic nucleus (SCN) is the central circadian pacemaker that governs rhythmic fluctuations in behavior and physiology in a 24-hr cycle and synchronizes them to the external environment by daily resetting in response to light. The bilateral SCN is comprised of a mere ~20,000 neurons serving as cellular oscillators, a fact that has, until now, hindered the systematic study of the SCN on a global proteome level. Here we developed a fully automated and integrated proteomics platform, termed AutoProteome system, for an in-depth analysis of the light-responsive proteome of the murine SCN. All requisite steps for a large-scale proteomic study, including preconcentration, buffer exchanging, reduction, alkylation, digestion and online two-dimensional liquid chromatography-tandem MS analysis, are performed automatically on a standard liquid chromatography-MS system. As low as 2 ng of model protein bovine serum albumin and up to 20 μg and 200 μg of SCN proteins can be readily processed and analyzed by this system. From the SCN tissue of a single mouse, we were able to confidently identify 2131 proteins, of which 387 were light-regulated based on a spectral counts quantification approach. Bioinformatics analysis of the light-inducible proteins reveals their diverse distribution in different canonical pathways and their heavy connection in 19 protein interaction networks. The AutoProteome system identified vasopressin-neurophysin 2-copeptin and casein kinase 1 delta, both of which had been previously implicated in clock timing processes, as light-inducible proteins in the SCN. Ras-specific guanine nucleotide-releasing factor 1, ubiquitin protein ligase E3A, and X-linked ubiquitin specific protease 9, none of which had previously been implicated in SCN clock timing processes, were also identified in this study as light-inducible proteins. The AutoProteome system opens a new avenue to systematically explore the proteome-wide events that occur in the SCN, either in response to light or other stimuli, or as a consequence of its intrinsic pacemaker capacity. |
Authors:
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Ruijun Tian; Matias Alvarez-Saavedra; Hai-Ying M Cheng; Daniel Figeys |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2011-08-22 |
Journal Detail:
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Title: Molecular & cellular proteomics : MCP Volume: 10 ISSN: 1535-9484 ISO Abbreviation: Mol. Cell Proteomics Publication Date: 2011 Nov |
Date Detail:
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Created Date: 2011-11-04 Completed Date: 2012-03-05 Revised Date: 2013-06-27 |
Medline Journal Info:
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Nlm Unique ID: 101125647 Medline TA: Mol Cell Proteomics Country: United States |
Other Details:
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Languages: eng Pagination: M110.007252 Citation Subset: IM |
Affiliation:
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Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Ontario, Canada K1H 8M5. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Automation, Laboratory* Cattle Chromatography, Liquid / standards Circadian Clocks* Gene Expression / radiation effects Light* Male Metabolic Networks and Pathways Mice Mice, Inbred C57BL Protein Interaction Maps Proteome / genetics, isolation & purification, metabolism* Serum Albumin, Bovine / standards Suprachiasmatic Nucleus / metabolism*, radiation effects Tandem Mass Spectrometry / standards |
| Grant Support | |
ID/Acronym/Agency:
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086549//Canadian Institutes of Health Research |
| Chemical | |
Reg. No./Substance:
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0/Proteome; 0/Serum Albumin, Bovine |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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