Backbone dynamics of oxidised and reduced forms of human atrial natriuretic peptide.
The backbone dynamics of the 28 residue 15N-labelled human atrial natriuretic peptide have been examined by 15N NMR methods. 15N R1, R2 and [1H]-15N NOE values were determined for the oxidised and reduced forms of the peptide (ANPox and ANPrd, respectively), and analysed using reduced spectral density mapping and an extended model-free approach. The two forms possessed correlation times for overall molecular motion of 4.7 ns and were highly flexible, with substantial contributions to relaxation processes from internal motions on picosecond to nanosecond time scales. Reduction of the Cys7-Cys23 disulphide bond to form ANPrd produced a very dynamic linear peptide with a mean overall order parameter of 0.2; the intramolecular cross-link in ANPox increased this to a mean value of 0.4. A simple model for segmental backbone motion accounted for the R2 values of both species using only two variable parameters, indicating that relaxation is dominated by interactions with sites <7 residues distant in the covalent network and that changes in the conformation of the disulphide bond lead to significant chemical exchange broadening in ANPox. The contributions of backbone dynamics to configurational entropy were determined and accounted for the different receptor binding affinities of cyclised and linear natriuretic peptides.
Peto
Heather
H
Centre for Protein Engineering, MRC Centre, Hills Road Cambridge CB2 2QH, UK.
Stott
Katherine
K
Sunde
Margaret
M
Broadhurst
R William
RW
D009320:Atrial Natriuretic Factor / Q000737:chemistry*; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D019277:Entropy; D004926:Escherichia coli / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D015233:Models, Statistical; D010100:Oxygen / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D013816:Thermodynamics
D004220:Disulfides; D008024:Ligands; D009320:Atrial Natriuretic Factor; D005982:Glutathione Transferase; D003545:Cysteine; D010100:Oxygen