Abstract
Receptor kinases play a central role in various biological processes, but due to their low abundance and highly hydrophobic and dynamic nature, only a few of them have been functionally characterized, and their partners and ligands remain unidentified. Receptor protein extraction and purification from plant tissues is one of the most challenging steps for the success of various biochemical analyses to characterize their function. Immunoprecipitation is a widely used and selective method for enriching or purifying a specific protein. Here we describe two different optimized protein purification protocols, batch and on-chip immunoprecipitation, which efficiently isolate plant membrane receptor kinases for functional analysis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Torii KU (2012) Mix-and-match: ligand-receptor pairs in stomatal development and beyond. Trends Plant Sci 17(12):711–719
Hazak O, Hardtke CS (2016) CLAVATA 1-type receptors in plant development. J Exp Bot 67(16):4827–4833
Wu Y, Zhou JM (2013) Receptor-like kinases in plant innate immunity. J Integr Plant Biol 55(12):1271–1286
Shpak ED (2013) Diverse roles of ERECTA family genes in plant development. J Integr Plant Biol 55(12):1238–1250
Niederhuth CE, Cho SK, Seitz K, Walker JC (2013) Letting go is never easy: abscission and receptor-like protein kinases. J Integr Plant Biol 55(12):1251–1263. doi:10.1111/jipb.12116
De Smet I, Voss U, Jurgens G, Beeckman T (2009) Receptor-like kinases shape the plant. Nat Cell Biol 11(10):1166–1173
Kim BH, Kim SY, Nam KH (2013) Assessing the diverse functions of BAK1 and its homologs in Arabidopsis, beyond BR signaling and PTI responses. Mol Cells 35(1):7–16
Kaboord B, Perr M (2008) Isolation of proteins and protein complexes by immunoprecipitation. Methods Mol Biol 424:349–364
van Zanten M, Snoek LB, Proveniers MC, Peeters AJ (2009) The many functions of ERECTA. Trends Plant Sci 14(4):214–218
Bemis SM, Lee JS, Shpak ED, Torii KU (2013) Regulation of floral patterning and organ identity by Arabidopsis ERECTA-family receptor kinase genes. J Exp Bot 64(17):5323–5333
Uchida N, Lee JS, Horst RJ, Lai HH, Kajita R, Kakimoto T, Tasaka M, Torii KU (2012) Regulation of inflorescence architecture by intertissue layer ligand-receptor communication between endodermis and phloem. Proc Natl Acad Sci U S A 109(16):6337–6342
Lee JS, Kuroha T, Hnilova M, Khatayevich D, Kanaoka MM, McAbee JM, Sarikaya M, Tamerler C, Torii KU (2012) Direct interaction of ligand-receptor pairs specifying stomatal patterning. Genes Dev 26(2):126–136
Lee JS, Hnilova M, Maes M, Lin YC, Putarjunan A, Han SK, Avila J, Torii KU (2015) Competitive binding of antagonistic peptides fine-tunes stomatal patterning. Nature 522(7557):439–443
Abrash EB, Bergmann DC (2010) Regional specification of stomatal production by the putative ligand CHALLAH. Development 137(3):447–455
Abrash EB, Davies KA, Bergmann DC (2011) Generation of signaling specificity in Arabidopsis by spatially restricted buffering of ligand-receptor interactions. Plant Cell 23(8):2864–2879
Takayama S, Shimosato H, Shiba H, Funato M, Che FS, Watanabe M, Iwano M, Isogai A (2001) Direct ligand-receptor complex interaction controls Brassica self-incompatibility. Nature 413(6855):534–538
Shpak ED, Lakeman MB, Torii KU (2003) Dominant-negative receptor uncovers redundancy in the Arabidopsis ERECTA Leucine-rich repeat receptor-like kinase signaling pathway that regulates organ shape. Plant Cell 15(5):1095–1110
Ogawa M, Shinohara H, Sakagami Y, Matsubayashi Y (2008) Arabidopsis CLV3 peptide directly binds CLV1 ectodomain. Science 319(5861):294
Shinohara H, Ogawa M, Sakagami Y, Matsubayashi Y (2007) Identification of ligand binding site of phytosulfokine receptor by on-column photoaffinity labeling. J Biol Chem 282(1):124–131
Bleckmann A, Weidtkamp-Peters S, Seidel CA, Simon R (2010) Stem cell signaling in Arabidopsis requires CRN to localize CLV2 to the plasma membrane. Plant Physiol 152(1):166–176
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Lee, J.S. (2017). Purification of Plant Receptor Kinases from Plant Plasma Membranes. In: Aalen, R. (eds) Plant Receptor Kinases. Methods in Molecular Biology, vol 1621. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7063-6_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7063-6_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7062-9
Online ISBN: 978-1-4939-7063-6
eBook Packages: Springer Protocols