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Integrating global proteomic and genomic expression profiles generated from islet α cells : Opportunities and challenges to deriving reliable biological inferences

Maziarz, Marlena LU ; Chung, Clement ; Drucker, Daniel J. and Emili, Andrew (2005) In Molecular and Cellular Proteomics 4(4). p.458-474
Abstract

Systematic profiling of expressed gene products represents a promising research strategy for elucidating the molecular phenotypes of islet cells. To this end, we have combined complementary genomic and proteomic methods to better assess the molecular composition of murine pancreatic islet glucagon-producing αTC-1 cells as a model system, with the expectation of bypassing limitations inherent to either technology alone. Gene expression was measured with an Affymetrix MG_U74Av2 oligonucleotide array, while protein expression was examined by performing high-resolution gel-free shotgun MS/MS on a nuclear-enriched cell extract. Both analyses were carried out in triplicate to control for experimental variability. Using a stringent detection p... (More)

Systematic profiling of expressed gene products represents a promising research strategy for elucidating the molecular phenotypes of islet cells. To this end, we have combined complementary genomic and proteomic methods to better assess the molecular composition of murine pancreatic islet glucagon-producing αTC-1 cells as a model system, with the expectation of bypassing limitations inherent to either technology alone. Gene expression was measured with an Affymetrix MG_U74Av2 oligonucleotide array, while protein expression was examined by performing high-resolution gel-free shotgun MS/MS on a nuclear-enriched cell extract. Both analyses were carried out in triplicate to control for experimental variability. Using a stringent detection p value cutoff of 0.04, 48% of all potential mRNA transcripts were predicted to be expressed (probes classified as present in at least two of three replicates), while 1,651 proteins were identified with high-confidence using rigorous database searching. Although 762 of 888 cross-referenced cognate mRNA-protein pairs were jointly detected by both platforms, a sizeable number (126) of gene products was detected exclusively by MS alone. Conversely, marginal protein identifications often had convincing microarray support. Based on these findings, we present an operational framework for both interpreting and integrating dual genomic and proteomic datasets so as to obtain a more reliable perspective into islet α cell function.

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author
; ; and
publishing date
type
Contribution to journal
publication status
published
in
Molecular and Cellular Proteomics
volume
4
issue
4
pages
458 - 474
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • scopus:17844369439
  • pmid:15741311
ISSN
1535-9476
DOI
10.1074/mcp.R500011-MCP200
language
English
LU publication?
no
id
c77c27ba-c329-4d0d-bb2a-1238a239deac
date added to LUP
2019-08-05 13:19:28
date last changed
2024-01-01 17:12:19
@article{c77c27ba-c329-4d0d-bb2a-1238a239deac,
  abstract     = {{<p>Systematic profiling of expressed gene products represents a promising research strategy for elucidating the molecular phenotypes of islet cells. To this end, we have combined complementary genomic and proteomic methods to better assess the molecular composition of murine pancreatic islet glucagon-producing αTC-1 cells as a model system, with the expectation of bypassing limitations inherent to either technology alone. Gene expression was measured with an Affymetrix MG_U74Av2 oligonucleotide array, while protein expression was examined by performing high-resolution gel-free shotgun MS/MS on a nuclear-enriched cell extract. Both analyses were carried out in triplicate to control for experimental variability. Using a stringent detection p value cutoff of 0.04, 48% of all potential mRNA transcripts were predicted to be expressed (probes classified as present in at least two of three replicates), while 1,651 proteins were identified with high-confidence using rigorous database searching. Although 762 of 888 cross-referenced cognate mRNA-protein pairs were jointly detected by both platforms, a sizeable number (126) of gene products was detected exclusively by MS alone. Conversely, marginal protein identifications often had convincing microarray support. Based on these findings, we present an operational framework for both interpreting and integrating dual genomic and proteomic datasets so as to obtain a more reliable perspective into islet α cell function.</p>}},
  author       = {{Maziarz, Marlena and Chung, Clement and Drucker, Daniel J. and Emili, Andrew}},
  issn         = {{1535-9476}},
  language     = {{eng}},
  month        = {{04}},
  number       = {{4}},
  pages        = {{458--474}},
  publisher    = {{American Society for Biochemistry and Molecular Biology}},
  series       = {{Molecular and Cellular Proteomics}},
  title        = {{Integrating global proteomic and genomic expression profiles generated from islet α cells : Opportunities and challenges to deriving reliable biological inferences}},
  url          = {{http://dx.doi.org/10.1074/mcp.R500011-MCP200}},
  doi          = {{10.1074/mcp.R500011-MCP200}},
  volume       = {{4}},
  year         = {{2005}},
}