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.
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX13QZ, UK.
Iain J
IJ
Day
Kiminori
K
Maeda
Howard J
HJ
Paisley
K Hun
KH
Mok
P J
PJ
Hore
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