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Identification of O-linked β-d-N-acetylglucosamine-Modified Proteins from Arabidopsis

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Plant Signalling Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 876))

Abstract

The posttranslational modification of proteins with O-linked β-d-N-acetylglucosamine (O-GlcNAc) on serine and threonine residues occurs in all animals and plants. This modification is dynamic and ubiquitous, and regulates many cellular processes, including transcription, signaling and cytokinesis and is associated with several diseases. Cycling of O-GlcNAc is tightly regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Plants have two OGTs, SPINDLY (SPY) and SECRET AGENT (SEC); disruption of both causes embryo lethality. Despite O-GlcNAc modification of proteins being discovered more than 20-years ago, identification and mapping of protein GlcNAcylation is still a challenging task. Here we describe the use of lectin affinity chromatography combined with electron transfer dissociation mass spectrometry to enrich and to detect O-GlcNAc modified peptides from Arabidopsis.

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Acknowledgments

This work was financially supported by the Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the US Department of Energy through Grant DE-FG02-08ER15973, and by NIH (R01GM066258) and NSF (IOS-0724688). The UCSF Mass Spectrometry Facility (A.L.B., Director) is supported by the Biomedical Research Technology Program of the National Centre for Research Resources, NIH NCRR RR01614.

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Authors and Affiliations

  1. Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA

    Shou-Ling Xu & Zhi-Yong Wang

  2. Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, USA

    Robert J. Chalkley & Alma L. Burlingame

Authors
  1. Shou-Ling Xu
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  2. Robert J. Chalkley
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  3. Zhi-Yong Wang
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  4. Alma L. Burlingame
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Corresponding author

Correspondence to Alma L. Burlingame .

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Editors and Affiliations

  1. Dept. Plant Biology, Carnegie Institution, Panama Street 260, Stanford, 94305, California, USA

    Zhi-Yong Wang

  2. Dept. Botany & Plant Science, University of California, Riverside, University Avenue 900, Riverside, 92521, California, USA

    Zhenbiao Yang

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Xu, SL., Chalkley, R.J., Wang, ZY., Burlingame, A.L. (2011). Identification of O-linked β-d-N-acetylglucosamine-Modified Proteins from Arabidopsis . In: Wang, ZY., Yang, Z. (eds) Plant Signalling Networks. Methods in Molecular Biology, vol 876. Humana Press. https://doi.org/10.1007/978-1-61779-809-2_3

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  • DOI: https://doi.org/10.1007/978-1-61779-809-2_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-808-5

  • Online ISBN: 978-1-61779-809-2

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