Methods to study protein folding by stopped-flow FT-IR.
Stopped-flow mixing coupled with time-resolved Fourier transform infrared (FT-IR) spectroscopy represents a new experimental approach to explore protein folding events, which has become possible only recently with the development of appropriate techniques. Here, we discuss experimental apparatus that are capable of initiating and monitoring protein folding processes on the millisecond to minute timescale. The strongest point of the FT-IR approach as a structure-specific probe is that a complete spectrum is available for each time point of measurement. In this way, several spectral windows are accessible simultaneously for the observation of the unfolding or the formation of different secondary structure elements and also events that can be attributed to changes in tertiary structure. One specific advantage of the infrared technique is the ability to monitor directly the kinetics of processes involving beta-sheet structures, which is exceptionally difficult to do with other techniques.
Fabian
Heinz
H
Robert Koch-Institute, Biomedical Spectroscopy, P 13, Nordufer 20, D-13353 Berlin, Germany.
Naumann
Dieter
D
D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011328:Prions / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012996:Solutions / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013696:Temperature
D011328:Prions; D012996:Solutions; D012259:Ribonuclease, Pancreatic