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Metabolomic and proteomic analysis of a clonal insulin-producing beta-cell line (INS-1 832/13).

Fernandez, Celine LU ; Krus, Ulrika LU ; Hallgard, Elna; Spégel, Peter LU ; Holm, Cecilia LU ; Krogh, Morten LU ; Wårell, Kristofer LU ; James, Peter LU and Mulder, Hindrik LU (2008) In Journal of Proteome Research 7(1). p.400-411
Abstract
Metabolites generated from fuel metabolism in pancreatic beta-cells control exocytosis of insulin, a process which fails in type 2 diabetes. To identify and quantify these metabolites, global and unbiased analysis of cellular metabolism is required. To this end, polar metabolites, extracted from the clonal 832/13 beta-cell line cultured at 2.8 and 16.7 mM glucose for 48 h, were derivatized followed by identification and quantification, using gas chromatography (GC) and mass spectrometry (MS). After culture at 16.7 mM glucose for 48 h, 832/13 beta-cells exhibited a phenotype reminiscent of glucotoxicity with decreased content and secretion of insulin. The metabolomic analysis revealed alterations in the levels of 7 metabolites derived from... (More)
Metabolites generated from fuel metabolism in pancreatic beta-cells control exocytosis of insulin, a process which fails in type 2 diabetes. To identify and quantify these metabolites, global and unbiased analysis of cellular metabolism is required. To this end, polar metabolites, extracted from the clonal 832/13 beta-cell line cultured at 2.8 and 16.7 mM glucose for 48 h, were derivatized followed by identification and quantification, using gas chromatography (GC) and mass spectrometry (MS). After culture at 16.7 mM glucose for 48 h, 832/13 beta-cells exhibited a phenotype reminiscent of glucotoxicity with decreased content and secretion of insulin. The metabolomic analysis revealed alterations in the levels of 7 metabolites derived from glycolysis, the TCA cycle and pentose phosphate shunt, and 4 amino acids. Principal component analysis of the metabolite data showed two clusters, corresponding to the cells cultured at 2.8 and 16.7 mM glucose, respectively. Concurrent changes in protein expression were analyzed by 2-D gel electrophoresis followed by LC-MS/MS. The identities of 86 spots corresponding to 75 unique proteins that were significantly different in 832/13 beta-cells cultured at 16.7 mM glucose were established. Only 5 of these were found to be metabolic enzymes that could be involved in the metabolomic alterations observed. Anticipated changes in metabolite levels in cells exposed to increased glucose were observed, while changes in enzyme levels were much less profound. This suggests that substrate availability, allosteric regulation, and/or post-translational modifications are more important determinants of metabolite levels than enzyme expression at the protein level. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Proteome Research
volume
7
issue
1
pages
400 - 411
publisher
The American Chemical Society
external identifiers
  • pmid:18062666
  • wos:000252154200046
  • scopus:38649130988
ISSN
1535-3893
DOI
10.1021/pr070547d
language
English
LU publication?
yes
id
8825428a-5bd3-4ce3-a3e8-f7d144c0d827 (old id 1035575)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/18062666?dopt=Abstract
date added to LUP
2008-04-01 11:25:48
date last changed
2017-04-23 03:37:05
@article{8825428a-5bd3-4ce3-a3e8-f7d144c0d827,
  abstract     = {Metabolites generated from fuel metabolism in pancreatic beta-cells control exocytosis of insulin, a process which fails in type 2 diabetes. To identify and quantify these metabolites, global and unbiased analysis of cellular metabolism is required. To this end, polar metabolites, extracted from the clonal 832/13 beta-cell line cultured at 2.8 and 16.7 mM glucose for 48 h, were derivatized followed by identification and quantification, using gas chromatography (GC) and mass spectrometry (MS). After culture at 16.7 mM glucose for 48 h, 832/13 beta-cells exhibited a phenotype reminiscent of glucotoxicity with decreased content and secretion of insulin. The metabolomic analysis revealed alterations in the levels of 7 metabolites derived from glycolysis, the TCA cycle and pentose phosphate shunt, and 4 amino acids. Principal component analysis of the metabolite data showed two clusters, corresponding to the cells cultured at 2.8 and 16.7 mM glucose, respectively. Concurrent changes in protein expression were analyzed by 2-D gel electrophoresis followed by LC-MS/MS. The identities of 86 spots corresponding to 75 unique proteins that were significantly different in 832/13 beta-cells cultured at 16.7 mM glucose were established. Only 5 of these were found to be metabolic enzymes that could be involved in the metabolomic alterations observed. Anticipated changes in metabolite levels in cells exposed to increased glucose were observed, while changes in enzyme levels were much less profound. This suggests that substrate availability, allosteric regulation, and/or post-translational modifications are more important determinants of metabolite levels than enzyme expression at the protein level.},
  author       = {Fernandez, Celine and Krus, Ulrika and Hallgard, Elna and Spégel, Peter and Holm, Cecilia and Krogh, Morten and Wårell, Kristofer and James, Peter and Mulder, Hindrik},
  issn         = {1535-3893},
  language     = {eng},
  number       = {1},
  pages        = {400--411},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Metabolomic and proteomic analysis of a clonal insulin-producing beta-cell line (INS-1 832/13).},
  url          = {http://dx.doi.org/10.1021/pr070547d},
  volume       = {7},
  year         = {2008},
}