bi300222f_si_001.pdf (2.85 MB)
Disentangling the Coil: Modulation of Conformational and Dynamic Properties by Site-Directed Mutation in the Non-Native State of Hen Egg White Lysozyme
journal contribution
posted on 2012-04-24, 00:00 authored by Friederike Sziegat, Robert Silvers, Martin Hähnke, Malene Ringkjøbing Jensen, Martin Blackledge, Julia Wirmer-Bartoschek, Harald SchwalbeThe conformational analysis of non-native states of proteins
remains one of the most difficult problems in structural biology,
because such states are represented by a superimposition of several
states that are rapidly interconverting. Hence, model building of
the conformational ensemble remains challenging, although many different
biophysical observables can be determined in non-native states of
proteins. Here, we present a comprehensive analysis of non-native
states of wild-type and mutant forms of the model protein lysozyme
by nuclear magnetic resonance spectroscopy. Relaxation rates, chemical
shifts, backbone and side chain coupling constants, residual dipolar
couplings, diffusion rate constants, and small-angle scattering data
merged with computational approaches, such as flexible meccano and ASTEROIDS, allow the description of the non-native state of
hen egg white lysozyme in unprecedented detail.