Characterization of tools to detect and enrich human and mouse O-GlcNAcase

Jennifer A. Groves, Natasha E. Zachara

Research output: Contribution to journalArticlepeer-review

Abstract

O-linked ß-N-acetylglucosamine (O-GlcNAc) is an essential regulatory post-translational modification of thousands of nuclear, cytoplasmic and mitochondrial proteins. O-GlcNAc is dynamically added and removed from proteins by the O-GlcNAc transferase and the O-GlcNAcase (OGA), respectively. Dysregulation of O-GlcNAc-cycling is implicated in the etiology of numerous diseases including tumorigenesis, metabolic dysfunction and neurodegeneration. To facilitate studies focused on the role of O-GlcNAc and OGA in disease, we sought to identify commercially available antibodies that enable the enrichment of full-length OGA (fOGA) from lysates of mouse and human origin. Here, we report that antibodies from Abcam and Bethyl Laboratories can be used to immunoprecipitate OGA to near-saturation from human and mouse cell lysates. However, western blotting analysis indicates that both antibodies, as well as three noncommercially available antibodies (345, 346, 352), detect fOGA and numerous cross-reacting proteins. These nonspecific signals migrate similarly to fOGA and are detected robustly, suggesting that the use of appropriate controls is essential to avoid the misidentification of OGA.

Original languageEnglish (US)
Pages (from-to)791-795
Number of pages5
JournalGlycobiology
Volume27
Issue number9
DOIs
StatePublished - Sep 1 2017

Keywords

  • Immunoprecipitation
  • Interaction
  • Mgea5
  • OGT
  • Signaling

ASJC Scopus subject areas

  • Biochemistry

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