Assays to Investigate the N-Glycosylation State and Function of Plant Pattern Recognition Receptors
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.
Key wordsAsparagine-linked glycosylation Protein quality control Endoglycosidase H Tunicamycin MAPK activation Seedling growth arrest Anthocyanin inhibition
This work was supported by T32 GM007499 (to S.A.L.) and Yale University Elizabeth Brown Postdoctoral Fellowship (to T.C.).
- 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 17:523–534CrossRefGoogle 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 25:93–105Google Scholar