Time-resolved FTIR difference spectroscopy as tool for investigating refolding reactions of ribonuclease T1 synchronized with trans --> cis prolyl isomerization.
The structurally well-characterized enzyme ribonuclease T1 was used as a model protein to further evaluate time-resolved Fourier transform IR difference spectroscopy in conjunction with temperature-jump techniques as a useful detection technique for protein folding studies. Compared to the wild-type protein, it was confirmed that the lack of one cis-proline bond at position 55 of the S54G/P55N variant is sufficient to significantly simplify and accelerate the refolding process. This result was sustained by the characterization of the early refolding events that occurred within the experimental dead time.
Moritz
Ralf
R
Robert Koch-Institut, P34, Nordufer 20, 13353 Berlin, Germany. ralf.moritz@gmx.de
Reinstädler
Diane
D
Fabian
Heinz
H
Naumann
Dieter
D
D007700:Kinetics; D011392:Proline / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D006163:Ribonuclease T1 / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013696:Temperature
D011994:Recombinant Proteins; D011392:Proline; D006163:Ribonuclease T1