Refolding of ribonuclease A monitored by real-time photo-CIDNP NMR spectroscopy.
Photo-CIDNP NMR spectroscopy is a powerful method for investigating the solvent accessibility of histidine, tyrosine and tryptophan residues in a protein. When coupled to real-time NMR, this technique allows changes in the environments of these residues to be used as a probe of protein folding. In this paper we describe experiments performed to monitor the refolding of ribonuclease A following dilution from a high concentration of chemical denaturant. These experiments provide a good example of the utility of this technique which provides information that is difficult to obtain by other biophysical methods. Real-time photo-CIDNP measurements yield residue-specific kinetic data pertaining to the folding reaction, interpreted in terms of current knowledge of the folding of bovine pancreatic ribonuclease A.
Day
Iain J
IJ
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX13QZ, UK.
Maeda
Kiminori
K
Paisley
Howard J
HJ
Mok
K Hun
KH
Hore
P J
PJ
D000818:Animals; D002417:Cattle; D019791:Guanidine / Q000737:chemistry; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010777:Photochemistry / Q000379:methods*; D011489:Protein Denaturation; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D014508:Urea / Q000737:chemistry
D014508:Urea; D012259:Ribonuclease, Pancreatic; D019791:Guanidine