H. Molinari et al., PROBING PROTEIN-STRUCTURE BY SOLVENT PERTURBATION OF NMR-SPECTRA - THE SURFACE ACCESSIBILITY OF BOVINE PANCREATIC TRYPSIN-INHIBITOR, Biophysical journal, 73(1), 1997, pp. 382-396
In the absence oi specific interactions, the relative attenuation of p
rotein NMR signals due to added stable free radicals such as TEMPOL sh
ould reflect the solvent accessibility of the molecular surface, The q
uantitative correlation between observed attenuation and surface acces
sibility was investigated with a model system, i.e., the small protein
bovine pancreatic trypsin inhibitor. A detailed discussion is present
ed on the reliability and limits of the approach, and guidelines are p
rovided for data acquisition, treatment, and interpretation. The NMR-d
erived accessibilities are compared with those obtained from x-ray dif
fraction and molecular dynamics data. Although the time-averaged acces
sibilities from molecular dynamics are ideally suited to fit the NMR d
ata, better agreement was observed between the paramagnetic attenuatio
ns of the fingerprint cross-peaks of homonuclear proton spectra and th
e total NH and H-alpha accessibilities calculated from x-ray coordinat
es, than from time-averaged molecular dynamics simulations. In additio
n, the solvent perturbation response appears to be a promising approac
h for detecting the thermal conformational evolution of secondary stru
cture elements in proteins.