Heating of proteins as a means of improving crystallization: a successful case study on a highly amyloidogenic triple mutant of human transthyretin.
The use of high temperatures in the purification procedures of heat-stable proteins is a well established technique. Recently, rapid pre-heat treatment of protein samples prior to crystallization trials was described as a final polishing step to improve the diffraction properties of crystals [Pusey et al. (2005), Prog. Biophys. Mol. Biol. 88, 359-386]. The present study demonstrates that extended high-temperature incubation (328 K for 48 h) of the highly amyloidogenic transthyretin mutant TTR G53S/E54D/L55S successfully removes heterogeneities and allows the reproducible growth of well diffracting crystals. Heat treatment might be applied as an optimization method to other cases in which the protein/biomolecule fails to form diffracting crystals.
Umeå Centre for Molecular Pathogenesis, Umeå University, SE-901 87 Umeå, Sweden.
Anders
A
Karlsson
A Elisabeth
AE
Sauer-Eriksson
D000682:Amyloid / Q000096:biosynthesis* / Q000235:genetics; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D018740:Genetic Heterogeneity; D006358:Hot Temperature; D006801:Humans; D009154:Mutation; D011228:Prealbumin / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification
D000682:Amyloid; D011228:Prealbumin