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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1035575
- author
- 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
- organization
- publishing date
- 2008
- 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 (ACS)
- external identifiers
-
- pmid:18062666
- wos:000252154200046
- scopus:38649130988
- pmid:18062666
- 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
- 2016-04-01 12:12:22
- date last changed
- 2023-11-11 16:40:49
@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 (ACS)}},
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}},
doi = {{10.1021/pr070547d}},
volume = {{7}},
year = {{2008}},
}