Quantitative MS-Based Proteomics: Comparing the MCF-7 Cellular Response to Hypoxia and a 2-Oxoglutarate Analogue

Chembiochem. 2020 Jun 2;21(11):1647-1655. doi: 10.1002/cbic.201900719. Epub 2020 Mar 3.

Abstract

The hypoxia-inducible factors (HIFs) are key transcription factors in determining cellular responses involving alterations in protein levels in response to limited oxygen availability in animal cells. 2-Oxoglutarate-dependent oxygenases play key roles in regulating levels of HIF and its transcriptional activity. We describe MS-based proteomics studies in which we compared the results of subjecting human breast cancer MCF-7 cells to hypoxia or treating them with a cell-penetrating derivative (dimethyl N-oxalylglycine; DMOG) of the stable 2OG analogue N-oxalylglycine. The proteomic results are consistent with reported transcriptomic analyses and support the proposed key roles of 2OG-dependent HIF prolyl- and asparaginyl-hydroxylases in the hypoxic response. Differences between the data sets for hypoxia and DMOG might reflect context-dependent effects or HIF-independent effects of DMOG.

Keywords: 2-oxoglutarate; hydroxylases; hypoxia; hypoxia-inducible factors (HIFs); oxygenases; proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Dicarboxylic / pharmacology*
  • Atlases as Topic
  • Cell Hypoxia / genetics*
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Gene Ontology
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • MCF-7 Cells
  • Metabolic Networks and Pathways / genetics
  • Molecular Sequence Annotation
  • Oxygen / pharmacology*
  • Proteome / classification
  • Proteome / genetics*
  • Proteome / metabolism
  • Proteomics / methods
  • Transcriptome*

Substances

  • Amino Acids, Dicarboxylic
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Proteome
  • Oxygen
  • oxalylglycine