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Real‐time NMR studies on folding of mutants of barnase and chymotrypsin inhibitor 2
Author(s) -
Killick Thomas R,
Freund Stefan M.V,
Fersht Alan R
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00075-1
Subject(s) - barnase , chemistry , folding (dsp implementation) , protein folding , stopped flow , phi value analysis , downhill folding , mutant , tryptophan , kinetics , fluorescence , biophysics , chymotrypsin , crystallography , biochemistry , reaction rate constant , amino acid , enzyme , biology , ribonuclease , rna , physics , trypsin , quantum mechanics , electrical engineering , gene , engineering
The folding and unfolding of proteins is generally assumed to be so co‐operative that the overall process may be followed by a single probe, such as tryptophan fluorescence. Folding kinetics of three mutants of barnase and chymotrypsin inhibitor 2 (CI2) were studied by real‐time NMR. Rate constants for changes in individual residues during the unfolding or refolding of the mutants studied by real‐time NMR are all within experimental error of the overall process of folding/unfolding measured by stopped‐flow measurements of tryptophan fluorescence. Folding of these mutants is thus highly co‐operative. Changes in the tryptophan fluorescence give accurate measurements of the protein folding process.