Abstract
Plant receptor kinases play diverse signaling roles in disease resistance and plant development. They represent a large plant gene family with over 600 members in Arabidopsis thaliana. While the functions of several members of the receptor kinase family have now been elucidated, a great proportion still remains uncharacterized. The structural and functional characterization of such plant receptor kinases may entail biochemical approaches that require access to purified protein, which can be made possible through heterologous protein expression. This chapter describes a strategy for expressing plant receptor kinases in E. coli, a bacterial host that has successfully been used to express and purify certain plant receptor kinase domains, some of which were subsequently used for biochemical assays. As full-length receptor-like kinases may be difficult to express, it is suggested to clone and express domains separately, after having identified domain borders using bioinformatics tools. A detailed cloning protocol is provided, as well as advice for testing expression efficiency and handling of expressed protein ending up in inclusion bodies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McCarty DR, Chory J (2000) Conservation and innovation in plant signaling pathways. Cell 103(2):201–209. doi:10.1016/S0092-8674(00)00113-6
Becraft PW (2002) Receptor kinase signaling in plant development. Annu Rev Cell Dev Biol 18:163–192. doi:10.1146/annurev.cellbio.18.012502.083431
Shiu SH, Bleecker AB (2001) Plant receptor-like kinase gene family: diversity, function, and signaling. Sci STKE 2001(113):re22. doi:10.1126/stke.2001.113.re22
Osakabe Y, Yamaguchi-Shinozaki K, Shinozaki K, Tran L-SP (2013) Sensing the environment: key roles of membrane-localized kinases in plant perception and response to abiotic stress. J Exp Bot 64(2):445–458. doi:10.1093/jxb/ers354
Tichtinsky G, Vanoosthuyse V, Cock JM, Gaude T (2003) Making inroads into plant receptor kinase signalling pathways. Trends Plant Sci 8(5):231–237. doi:10.1016/S1360-1385(03)00062-1
Torii KU (2009) Transmembrane receptors in plants: receptor kinases and their ligands. In: Annual plant reviews volume 33: intracellular signaling in plants. Wiley-Blackwell, pp 1–29. doi:10.1002/9781444302387.ch1
Walker JC, Zhang R (1990) Relationship of a putative receptor protein kinase from maize to the S-locus glycoproteins of Brassica. Nature 345(6277):743–746. doi:10.1038/345743a0
Shiu S-H, Karlowski WM, Pan R, Tzeng Y-H, Mayer KFX, Li W-H (2004) Comparative analysis of the receptor-like kinase family in Arabidopsis and rice. Plant Cell 16(5):1220–1234. doi:10.1105/tpc.020834
Shiu S-H, Bleecker AB (2001) Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc Natl Acad Sci USA 98(19):10763–10768. doi:10.1073/pnas.181141598
Shiu SH, Bleecker AB (2003) Expansion of the receptor-like kinase/Pelle gene family and receptor-like proteins in Arabidopsis. Plant Physiol 132(2):530–543. doi:10.1104/pp.103.021964
Fischer I, Dievart A, Droc G, Dufayard JF, Chantret N (2016) Evolutionary dynamics of the leucine-rich repeat receptor-like kinase (LRR-RLK) subfamily in angiosperms. Plant Physiol 170(3):1595–1610. doi:10.1104/pp.15.01470
Gou X, He K, Yang H, Yuan T, Lin H, Clouse SD, Li J (2010) Genome-wide cloning and sequence analysis of leucine-rich repeat receptor-like protein kinase genes in Arabidopsis thaliana. BMC Genomics 11(1):1–15. doi:10.1186/1471-2164-11-19
Han Z, Sun Y, Chai J (2014) Structural insight into the activation of plant receptor kinases. Curr Opin Plant Biol 20:55–63. doi:10.1016/j.pbi.2014.04.008
Horn MA, Walker JC (1995) Chapter 37 Expression and assay of autophosphorylation of recombinant protein kinases. In: Galbraith DW, Bohnert HJ, Bourque PB (eds) Methods in cell biology, vol 49. Academic Press, pp 531–541. doi:10.1016/S0091-679X(08)61478-8
Peti W, Page R (2007) Strategies to maximize heterologous protein expression in Escherichia coli with minimal cost. Protein Expr Purif 51(1):1–10. doi:10.1016/j.pep.2006.06.024
Horn MA, Walker JC (1994) Biochemical properties of the autophosphorylation of RLK5, a receptor-like protein kinase from Arabidopsis thaliana. Biochim Biophys Acta 1208(1):65–74
Oh M-H, Ray WK, Huber SC, Asara JM, Gage DA, Clouse SD (2000) Recombinant brassinosteroid insensitive 1 receptor-like kinase autophosphorylates on serine and threonine residues and phosphorylates a conserved peptide motif in vitro. Plant Physiol 124(2):751–766
Li J, Wen J, Lease KA, Doke JT, Tax FE, Walker JC (2002) BAK1, an Arabidopsis LRR receptor-like protein kinase, interacts with BRI1 and modulates brassinosteroid signaling. Cell 110(2):213–222. doi:10.1016/S0092-8674(02)00812-7
Tameling WI, Elzinga SD, Darmin PS, Vossen JH, Takken FL, Haring MA, Cornelissen BJ (2002) The tomato R gene products I-2 and MI-1 are functional ATP binding proteins with ATPase activity. Plant Cell 14(11):2929–2939
Boyes DC, Nam J, Dangl JL (1998) The Arabidopsis thaliana RPM1 disease resistance gene product is a peripheral plasma membrane protein that is degraded coincident with the hypersensitive response. Proc Natl Acad Sci USA 95(26):15849–15854
Afzal AJ, Lightfoot DA (2007) Soybean disease resistance protein RHG1-LRR domain expressed, purified and refolded from Escherichia coli inclusion bodies: preparation for a functional analysis. Protein Expr Purif 53(2):346–355. doi:10.1016/j.pep.2006.12.017
Baneyx F, Mujacic M (2004) Recombinant protein folding and misfolding in Escherichia coli. Nat Biotechnol 22(11):1399–1408. doi:10.1038/nbt1029
Fischer B, Perry B, Sumner I, Goodenough P (1992) A novel sequential procedure to enhance the renaturation of recombinant protein from Escherichia coli inclusion bodies. Protein Eng 5(6):593–596
Misawa S, Kumagai I (1999) Refolding of therapeutic proteins produced in Escherichia coli as inclusion bodies. Biopolymers 51(4):297–307. doi:10.1002/(sici)1097-0282(1999)51:4<297::aid-bip5>3.0.co;2-i
Mitchell A, Chang H-Y, Daugherty L, Fraser M, Hunter S, Lopez R, McAnulla C, McMenamin C, Nuka G, Pesseat S, Sangrador-Vegas A, Scheremetjew M, Rato C, Yong S-Y, Bateman A, Punta M, Attwood TK, Sigrist CJA, Redaschi N, Rivoire C, Xenarios I, Kahn D, Guyot D, Bork P, Letunic I, Gough J, Oates M, Haft D, Huang H, Natale DA, Wu CH, Orengo C, Sillitoe I, Mi H, Thomas PD, Finn RD (2015) The InterPro protein families database: the classification resource after 15 years. Nucleic Acids Res 43(Database issue):D213–D221. doi:10.1093/nar/gku1243
Lobley A, Sadowski MI, Jones DT (2009) pGenTHREADER and pDomTHREADER: new methods for improved protein fold recognition and superfamily discrimination. Bioinformatics 25(14):1761–1767. doi:10.1093/bioinformatics/btp302
Structural Genomics C, Architecture et Fonction des Macromolécules B, Berkeley Structural Genomics C, China Structural Genomics C, Integrated Center for S, Function I, Israel Structural Proteomics C, Joint Center for Structural G, Midwest Center for Structural G, New York Structural Genomi XRCfSG, Northeast Structural Genomics C, Oxford Protein Production F, Protein Sample Production Facility MDCfMM, Initiative RSGP, Complexes S (2008) Protein production and purification. Nat Methods 5(2):135–146. doi:10.1038/nmeth.f.202
McGuffin LJ, Bryson K, Jones DT (2000) The PSIPRED protein structure prediction server. Bioinformatics 16(4):404–405. doi:10.1093/bioinformatics/16.4.404
Reich S, Puckey LH, Cheetham CL, Harris R, Ali AA, Bhattacharyya U, Maclagan K, Powell KA, Prodromou C, Pearl LH, Driscoll PC, Savva R (2006) Combinatorial domain hunting: an effective approach for the identification of soluble protein domains adaptable to high-throughput applications. Protein Sci 15(10):2356–2365. doi:10.1110/ps.062082606
Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5:172. doi:10.3389/fmicb.2014.00172
Costa S, Almeida A, Castro A, Domingues L (2014) Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system. Front Microbiol 5:63. doi:10.3389/fmicb.2014.00063
Kapust RB, Waugh DS (1999) Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused. Protein Sci 8(8):1668–1674. doi:10.1110/ps.8.8.1668
Terpe K (2003) Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 60(5):523–533. doi:10.1007/s00253-002-1158-6
Kimple ME, Brill AL, Pasker RL (2013) Overview of affinity tags for protein purification. Curr Protoc Protein Sci. doi:10.1002/0471140864.ps0909s73
Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and heterologous protein expression. Trends Biotechnol 22(7):346–353. doi:10.1016/j.tibtech.2004.04.006
Miroux B, Walker JE (1996) Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 260(3):289–298. doi:10.1006/jmbi.1996.0399
Korpimaki T, Kurittu J, Karp M (2003) Surprisingly fast disappearance of beta-lactam selection pressure in cultivation as detected with novel biosensing approaches. J Microbiol Methods 53(1):37–42
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
Agha, M.A., Lightfoot, D., Afzal, A.J. (2017). Expression of Plant Receptor Kinases in E. coli . 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_1
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7063-6_1
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