Skip to main content
SpringerLink
Log in
Book cover

Plant Pattern Recognition Receptors pp 61–79Cite as

Assays to Investigate the N-Glycosylation State and Function of Plant Pattern Recognition Receptors

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1578))

Abstract

The biogenesis and functionality of pattern recognition receptors (PRRs) are critical for robust plant immune responses. Here, we present methods to determine the N-glycosylation state and ligand-induced activity of these receptors for comparative quantitative analysis. These techniques can be used to identify mutants and chemical inhibitors affecting PRR biogenesis and functionality. When combined, these techniques can provide useful insights on biological processes necessary to synthesize a properly membrane-localized and ligand-responsive PRR.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Aebi M, Bernasconi R, Clerc S, Molinari M (2010) N-Glycan structures: recognition and processing in the ER. Trends Biochem Sci 35:74–82

    Article  CAS  PubMed  Google Scholar 

  2. Strasser R (2014) Biological significance of complex N-glycans in plants and their impact on plant physiology. Front Plant Sci 5:363

    Article  PubMed  PubMed Central  Google Scholar 

  3. Gómez-Gómez L, Boller T (2000) FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol Cell 5:1003–1011

    Article  PubMed  Google Scholar 

  4. Zipfel C, Kunze G, Chinchilla D, Caniard A, Jones JDG, Boller T, Felix G (2006) Perception of the bacterial PAMP EF-Tu by the receptor EFR restricts Agrobacterium-mediated transformation. Cell 125:749–760

    Article  CAS  PubMed  Google Scholar 

  5. Saijo Y (2010) ER quality control of immune receptors and regulators in plants. Cell Microbiol 12:716–724

    Article  CAS  PubMed  Google Scholar 

  6. Li J, Zhao-Hui C, Batoux M, Nekrasov V, Roux M, Chinchilla D et al (2009) Specific ER quality control components required for biogenesis of the plant innate immune receptor EFR. Proc Natl Acad Sci U S A 106:15973–15978

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Lu X, Tintor N, Mentzel T, Kombrink E, Boller T, Robatzek S et al (2009) Uncoupling of sustained MAMP receptor signaling from early outputs in an Arabidopsis endoplasmic reticulum glucosidease II allele. Proc Natl Acad Sci U S A 106:22522–22527

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Nekrasov V, Li J, Batoux M, Roux M, Chu Z-H, Lacombe S, Bougon A et al (2009) Control of the pattern-recognition receptor EFR by an ER protein complex in plant immunity. EMBO J 28:3428–3438

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Saijo Y, Tintor N, Lu X, Rauf P, Pajerowska-Mukhtar K, Häweker H et al (2009) Receptor quality control in the endoplasmic reticulum for plant innate immunity. EMBO J 28:3439–3449

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. von Numers N, Survila M, Aalto M, Batoux M, Heino P, Palva ET et al (2010) Requirement of a homolog of glucosidease II beta-subunit for EFR-mediated defense signaling in Arabidopsis thaliana. Mol Plant 3:740–750

    Article  Google Scholar 

  11. Häweker H, Rips S, Koiwa H, Salomon S, Saijo Y, Chinchilla D, Robatzek S, von Schaewen A (2010) Pattern recognition receptors require N-glycosylation to mediate plant immunity. J Biol Chem 285:4629–4636

    Article  PubMed  Google Scholar 

  12. Sun W, Cao Y, Jansen KL, Bittel P, Boller T, Bent AF (2012) Probing the Arabidopsis flagellin receptor: FLS2-FLS2 association and the contributions of specific domains to signaling function. Plant Cell 24:1096–1113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Chinchilla D, Bauer Z, Regenass M, Boller T, Felix G (2006) The Arabidopsis receptor kinase FLS2 binds flg22 and determines the specificity of flagellin perception. Plant Cell 18:465–476

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Elbein AD (1987) Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu Rev Biochem 56:497–534

    Article  CAS  PubMed  Google Scholar 

  15. Martínez IM, Chrispeels MJ (2003) Genomic analysis of the unfolded protein response in Arabidopsis shows its connection to important cellular processes. Plant Cell 15:561–576

    Article  PubMed  PubMed Central  Google Scholar 

  16. Iwata Y, Koizumi N (2005) An Arabidopsis transcription factor, AtbZIP60, regulates the endoplasmic reticulum stress response in a manner unique to plants. Proc Natl Acad Sci U S A 102:5280–5285

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Lozoya E, Block A, Lois R, Hahlbrock K, Scheel D (1991) Transcriptional repression of light-induced flavonoid synthesis by elicitor treatment of cultured parsley cells. Plant J 1:277–234

    Article  Google Scholar 

  18. Lo SC, Nicholson RL (1998) Reduction of light-induced anthocyanin accumulation in inoculated sorghum mesocotyls. Implications for a compensatory role in the defense response. Plant Physiol 116:979–989

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. McLusky SR, Bennett MH, Beale MH, Lewis MJ, Gaskin P, Mansfield JW (1999) Cell wall alterations and localized accumulation of feruloyl-3’-methoxytyramine in onion epidermis at sites of attempted penetration by Botrytis allii are associated with actin polarisation, peroxidase activity and suppression of flavonoid biosynthesis. Plant J 17:523–534

    Article  CAS  Google Scholar 

  20. Serrano M, Kanehara K, Torres M, Yamada K, Tintor N, Kombrink E et al (2012) Repression of sucrose/ultraviolet B light-induced flavonoid accumulation in microbe-associated molecular pattern-triggered immunity in Arabidopsis. Plant Physiol 158:408–422

    Article  CAS  PubMed  Google Scholar 

  21. Korasick DA, McMichael C, Walker KA, Anderson JC, Bednarek SY (2010) Novel functions of Stomatal Cytokinesis-Defective 1 (SCD1) in innate immune responses against bacteria. J Biol Chem 285:23342–23350

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Smith JM, Leslie ME, Robinson SJ, Korasick DA, Zhang T, Backues SK et al (2014) Loss of Arabidopsis thaliana dynamin-related protein 2B reveals separation of innate immune signaling pathways. PLoS Pathog 10:e1004578

    Article  PubMed  PubMed Central  Google Scholar 

  23. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  24. Mancinelli AL, Schwartz OM (1984) The photoregulation of the anthocyanin synthesis IX. The photosensitivity of the response in dark and light-grown tomato seedlings. Plant Cell Physiol 25:93–105

    CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by T32 GM007499 (to S.A.L.) and Yale University Elizabeth Brown Postdoctoral Fellowship (to T.C.).

Author information

Authors and Affiliations

  1. Department of Molecular, Cellular & Developmental Biology, Yale University, 266 Whitney Ave, New Haven, CT, 06510, USA

    Stacey A. Lawrence, Teresa Ceserani & Nicole K. Clay

Authors
  1. Stacey A. Lawrence
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Teresa Ceserani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nicole K. Clay
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nicole K. Clay .

Editor information

Editors and Affiliations

  1. Department of Plant Pathology and Microbiology Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Libo Shan

  2. Department of Biochemistry and Biophysics Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Ping He

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Lawrence, S.A., Ceserani, T., Clay, N.K. (2017). Assays to Investigate the N-Glycosylation State and Function of Plant Pattern Recognition Receptors. In: Shan, L., He, P. (eds) Plant Pattern Recognition Receptors. Methods in Molecular Biology, vol 1578. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6859-6_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6859-6_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6858-9

  • Online ISBN: 978-1-4939-6859-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation