Characterisation of urea-denatured states of an immunoglobulin superfamily domain by heteronuclear NMR.

The structural and dynamic properties of an immunoglobulin superfamily domain (IgSF), Ig 18', have been characterised by NMR at 285 K, in the presence of 4.2 M and 6.0 M urea, respectively. Analysis of chemical shift deviations, 3JHNHalpha coupling constants, sequential NOE pattern, and 15N relaxation data reveals that although the two urea-denatured states are highly disordered, some local turn-like residual structures do exist. Moreover, some distinct differences between the properties of the two denatured states are observed. In 4.2 M urea-denatured Ig 18', regions 80-83 and 86-92 adopt turn-like conformations, furthermore, region 84-93 is involved in slow exchange processes that occur on a micro- to millisecond time-scale. In the 6.0 M urea-denatured state, these turn-like conformations are less occupied, and chemical exchange processes in region 84-93 are largely reduced. In contrast, region 32-36 has persistent turn-like structures in both urea-denatured states. Some correlation between the spectral density function at 0 frequency, Jeff(0), for the urea-denatured states and the secondary structure elements of the folded state have been observed. Except for the terminal regions, residues corresponding to beta-strands have higher Jeff(0) values compared to residues corresponding to loops. The characterisation and comparison of the two urea-denatured states highlight residues that possess properties that may be crucial for the initiation of folding of this domain. Copyright 1998 Academic Press Limited.