Temperature-jump-induced refolding of ribonuclease A: a time-resolved FTIR spectroscopic study.
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 degrees C into a special cuvette system kept at 20 degrees 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 beta-structures and the disappearance of irregular conformations could be observed in this time interval.
Backmann
J
J
Robert Koch-Institut, Fachgebiet 233 'IR-Spektroskopie und Schnelldiagnostik', Berlin, Germany.
Fabian
H
H
Naumann
D
D
D000818:Animals; D002417:Cattle; D066298:In Vitro Techniques; D007700:Kinetics; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation; D013696:Temperature; D013816:Thermodynamics
D012259:Ribonuclease, Pancreatic