Abstract
Src homology 2 (SH2) domains are key modulators in various signaling pathways allowing the recognition of phosphotyrosine sites of different proteins. Despite the fact that SH2 domains acquire their biological functions in a monomeric state, a multitude of reports have shown their tendency to dimerize. Here, we provide a technical description on how to isolate and characterize by gel filtration, circular dichroism (CD), and nuclear magnetic resonance (NMR) each conformational state of p59fyn SH2 domain.
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References
Kaneko T, Joshi R, Feller SM, Li SS (2012) Phosphotyrosine recognition domains: the typical, the atypical and the versatile. Cell Commun Signal 10(1):32. doi:10.1186/1478-811X-10-32
Huculeci R, Garcia-Pino A, Buts L, Lenaerts T, van Nuland N (2015) Structural insights into the intertwined dimer of fyn SH2. Protein Sci. doi:10.1002/pro.2806
Mackinnon SS, Malevanets A, Wodak SJ (2013) Intertwined associations in structures of homooligomeric proteins. Structure 21(4):638–649. doi:10.1016/j.str.2013.01.019
Maurer-Stroh S, Debulpaep M, Kuemmerer N, Lopez de la Paz M, Martins IC, Reumers J, Morris KL, Copland A, Serpell L, Serrano L, Schymkowitz JW, Rousseau F (2010) Exploring the sequence determinants of amyloid structure using position-specific scoring matrices. Nat Methods 7(3):237–242. doi:10.1038/nmeth.1432
Schiering N, Casale E, Caccia P, Giordano P, Battistini C (2000) Dimer formation through domain swapping in the crystal structure of the Grb2-SH2-Ac-pYVNV complex. Biochemistry 39(44):13376–13382
Rane CK, Minden A (2014) P21 activated kinases: structure, regulation, and functions. Small GTPases 5:28003. doi:10.4161/sgtp.28003
Sambrook J, Fritsch EF, Maniatis F (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, NY
Farrow NA, Muhandiram R, Singer AU, Pascal SM, Kay CM, Gish G, Shoelson SE, Pawson T, Forman-Kay JD, Kay LE (1994) Backbone dynamics of a free and phosphopeptide-complexed Src homology 2 domain studied by 15N NMR relaxation. Biochemistry 33(19):5984–6003
Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6(3):277–293
Vranken WF, Boucher W, Stevens TJ, Fogh RH, Pajon A, Llinas M, Ulrich EL, Markley JL, Ionides J, Laue ED (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59(4):687–696. doi:10.1002/prot.20449
Acknowledgments
R.H., F.K., Nv.N., L.B., A.G., and T.L. were supported by Fonds Wetenschappelijk Onderzoek (FWO) via research grants G.0.116.09.N.10 and G025915N. Additional funding for T.L. was provided by Fondation de la Recherche Scientifique (FRS-FNRS) via the FRFC grant 2.4606.11. The VIB and the Hercules Foundation support the research in Nv.N. group.
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Huculeci, R., Kieken, F., Garcia-Pino, A., Buts, L., van Nuland, N., Lenaerts, T. (2017). Structural Characterization of Monomeric/Dimeric State of p59fyn SH2 Domain. In: Machida, K., Liu, B. (eds) SH2 Domains. Methods in Molecular Biology, vol 1555. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6762-9_14
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DOI: https://doi.org/10.1007/978-1-4939-6762-9_14
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