Abstract
Construct design and sample formulation are critical in structural biology projects. Large numbers of sample variants are often produced and analyzed for a single target and significant effort is dedicated to sample characterization in order to identify at an early stage the most promising samples to help save manpower and time. Here, we present a method based on a thermal stability assay that can help estimate the likelihood of biological samples to produce crystals. This assay is rapid, inexpensive and consumes very small amounts of sample. The results can be used to prioritize certain constructs at an early stage or as an objective test to help decide when to undertake other type of approaches addressed at improving sample properties.
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References
Zulauf M, D'Arcy A (1992) J Cryst Growth 122:102–106
Banci L, Bertini I, Cusack S et al (2006) Acta Crystallogr D 62:1208–1217
Graslund S, Nordlund P, Weigelt J et al (2008) Nat Methods 5:135–146
Geerlof A, Brown J, Coutard B et al (2006) Acta Crystallogr D 62:1125–1136
Niesen FH, Berglund H, Vedadi M (2007) Nat Protoc 2:2212–2221
Ericsson UB, Hallberg BM, Detitta GT et al (2006) Anal Biochem 357:289–298
Vedadi M, Niesen FH, Allali-Hassani A et al (2006) Proc Natl Acad Sci USA 103: 15835–15840
Pantoliano MW, Petrella EC, Kwasnoski JD et al (2001) J Biomol Screen 6:429–440
Hawe A, Sutter M, Jiskoot W (2008) Pharm Res 25:1487–1499
Dupeux F, Rower M, Seroul G et al (2011) Acta Crystallogr D 67:915–919
Matulis D, Kranz JK, Salemme FR et al (2005) Biochemistry 44:5258–5266
Heads JT, Adams R, D'Hooghe LE et al (2012) Protein Sci 21:1315–1322
Forneris F, Orru R, Bonivento D et al (2009) FEBS J 276:2833–2840
Alexandrov AI, Mileni M, Chien EY et al (2008) Structure 16:351–359
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Mariaule, V., Dupeux, F., Márquez, J.A. (2014). Estimation of Crystallization Likelihood Through a Fluorimetric Thermal Stability Assay. In: Chen, Y. (eds) Structural Genomics. Methods in Molecular Biology, vol 1091. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-691-7_13
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DOI: https://doi.org/10.1007/978-1-62703-691-7_13
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-690-0
Online ISBN: 978-1-62703-691-7
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