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Temperature‐jump‐induced refolding of ribonuclease A: A time‐resolved FTIR spectroscopic study
Author(s) -
Jan Backmann,
Heinz Fabian,
Dieter Naumann
Publication year - 1995
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 - 10.1016/0014-5793(95)00387-o
Subject(s) - cuvette , fourier transform infrared spectroscopy , chemistry , temperature jump , ribonuclease , bovine pancreatic ribonuclease , rnase p , kinetics , folding (dsp implementation) , spectroscopy , analytical chemistry (journal) , crystallography , chromatography , optics , physics , biochemistry , rna , electrical engineering , quantum mechanics , gene , engineering
FTIR difference spectroscopy has been used for the first time to investigate the kinetics of secondary structure formation during refolding. The refolding process of ribonuclease A (RNase A) as a model system was induced by applying a temperature‐jump of 60 degrees. The temperature‐jump was triggered by rapidly injecting a small volume of the thermally unfolded protein solution at 80°C into a special cuvette system kept at 20°C. The dead‐time of the injection and the time resolution of the FTIR spectrometer permitted the observation of refolding processes in a time window ranging from 170 ms to several minutes. Specifically, the formation of β‐structures and the disappearance of irregular conformations could be observed in this time interval.