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Comparative proteomic analysis of the insulin-induced L6 myotube secretome

Author(s)
Yoon, Jong HyukYea, KyungmooKim, JaeyoonChoi, Yoon SupPark, SehoonLee, HyeongjiLee, Chang SupSuh, Pann-GhillRyu, Sung Ho
Issued Date
2009-01
DOI
10.1002/pmic.200800187
URI
https://scholarworks.unist.ac.kr/handle/201301/5652
Fulltext
http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=59449096103
Citation
PROTEOMICS, v.9, no.1, pp.51 - 60
Abstract
Emerging evidence has revealed an endocrine function for skeletal muscle; in fact, certain antiinflammatory cytokines are secreted only from contractile skeletal muscle. However, the skeletal muscle secretome as a whole is poorly characterized, as is how it changes in response to extracellular stimuli. Herein, we sought to identify and characterize the members of the skeletal muscle secretome, and to determine which protein secretion levels were modulated in response to insulin stimulation. To conduct these studies, we treated differentiated L6 rat skeletal muscle cells with insulin or left them untreated, and we comparatively analyzed the proteins secreted into the media. We fractionated this conditioned media using offline RP HPLC, digested the fractionated proteins, and analyzed the resulting peptides with LC-ESI-MS/MS. We identified a total of 254 proteins, and by using three different filtering methods, we identified 153 of these as secretory proteins. Fourteen proteins were secreted at higher levels under insulin stimulation, including several proteins known to be highly secreted in metabolic diseases; 19 proteins were secreted at lower levels under insulin stimulation. These result not only pinpointed several previously unknown, insulin induced, secretory proteins of skeletal muscle, it also described a novel approach for conditioned secretome analysis.
Publisher
WILEY-BLACKWELL
ISSN
1615-9853
Keyword (Author)
Liquid chromatography-tandem mass spectrometrySecreted proteinShotgun proteomics
Keyword
PLASMINOGEN-ACTIVATOR INHIBITOR-1SKELETAL-MUSCLEMATRIX METALLOPROTEINASESADIPOSE-TISSUEDIABETES-MELLITUSIGF-IEXPRESSIONGLUCOSECELLSPROTEINS

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