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Atomic mutations in annexin V. Thermodynamic studies of isomorphous protein variants
Citation key 010.1998.budisa
Author Budisa, N. and Huber, R. and Minks, C. and Golbik, R. and Weyher, E. and Moroder, L.
Pages 1-9
Year 1998
DOI 10.1046/j.1432-1327.1998.2530001.x
Journal Eur. J. Biochem.
Volume 253
Abstract We have recently developed methods for specific and high-level replacement of methionine with 2-aminohexanoic acid, selenomethionine and telluromethionine as isosteric and atomic analogues for structural investigations of human recombinant annexin V. The variants formed isomorphic crystals and retained the parent three-dimensional fold and bioactivities. Folding parameters were determined from thermal and chemical unfolding to partially denatured states. Stabilities estimated from guanidinium chloride unfolding equilibria are not changed significantly for the atomic mutants (S–>Se–>Te) while the denaturation midpoint is shifted toward lower values with an increase of the m values at the increase of hydrophobicity. In contrast, stabilities in urea are considerably affected by the atomic substitutions, decreasing together with the m and [D]1/2 values. The methylene and selenium variants are identical within the limits of error of all measurements performed here. The physical parameters of the amino acid analogues and the values derived from the slopes of the unfolding data are highly correlated. This approach demonstrates how systematic variations in atomic number at the site of replacement (atomic mutations) can provide a method to probe specific folding properties of proteins.
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