Document Detail

Uncovering the proteome response of the master circadian clock to light using an AutoProteome system.
MedLine Citation:
PMID:  21859948     Owner:  NLM     Status:  MEDLINE    
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.
Ruijun Tian; Matias Alvarez-Saavedra; Hai-Ying M Cheng; Daniel Figeys
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-08-22
Journal Detail:
Title:  Molecular & cellular proteomics : MCP     Volume:  10     ISSN:  1535-9484     ISO Abbreviation:  Mol. Cell Proteomics     Publication Date:  2011 Nov 
Date Detail:
Created Date:  2011-11-04     Completed Date:  2012-03-05     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  101125647     Medline TA:  Mol Cell Proteomics     Country:  United States    
Other Details:
Languages:  eng     Pagination:  M110.007252     Citation Subset:  IM    
Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Ontario, Canada K1H 8M5.
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MeSH Terms
Automation, Laboratory*
Chromatography, Liquid / standards
Circadian Clocks*
Gene Expression / radiation effects
Metabolic Networks and Pathways
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
086549//Canadian Institutes of Health Research
Reg. No./Substance:
0/Proteome; 0/Serum Albumin, Bovine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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