title: Real-time NMR monitoring of protein-folding kinetics by a recycle flow system for temperature jump.
abstract: An NMR method was developed that allows for real-time monitoring of reactions (on the order of seconds) induced by a temperature jump. In a recycle flow system, heating and cooling baths were integrated, with the latter inside the NMR probe. A refolding reaction of ribonuclease A was triggered by rapid cooling and monitored by a series of NMR measurements over 12 s. Data were processed by principal component analysis, in which a factor related to the structural change with an exponential rate constant of 0.2-0.7 s(-1) was successfully separated from factors related to baseline instability and/or noise. Temperature dependency of the rate constant revealed the entropy-driven formation of the transition state of the refolding reaction.
authors.affiliation: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) , 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Yamasaki
authors.forename: Yuji
authors.initials: Y
authors.lastname: Obara
authors.forename: Manabu
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamasaki
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Wakuda
authors.forename: Michiya
authors.initials: M
authors.lastname: Okada
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Kohzuma
mesh_terms: D000818:Animals; D002417:Cattle; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D025341:Principal Component Analysis; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D013696:Temperature; D013997:Time Factors
chemical_list: D012259:Ribonuclease, Pancreatic