title: Detection and characterization of a folding intermediate in barnase by NMR.
abstract: Protein engineering is being developed for mapping the energetics and pathway of protein folding. From kinetic studies on wild-type and mutant proteins, the sequence and energetics of formation of tertiary interactions of side chains can be mapped and the formation of secondary structure inferred. Here we cross-check and complement results from this approach by using a recently developed procedure that traps and characterizes secondary structure in intermediate states using 1H NMR. The refolding of barnase is shown to be a multiphasic process in which the secondary structure in alpha-helices and beta-sheets and some turns is formed more rapidly than is the overall folding.
authors.affiliation: Department of Chemistry, University of Cambridge, UK.
authors.forename: M
authors.initials: M
authors.lastname: Bycroft
authors.forename: A
authors.initials: A
authors.lastname: Matouschek
authors.forename: J T
authors.initials: JT
authors.lastname: Kellis
authors.forename: L
authors.initials: L
authors.lastname: Serrano
authors.forename: A R
authors.initials: AR
authors.lastname: Fersht
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D006860:Hydrogen Bonding; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011489:Protein Denaturation; D012260:Ribonucleases
chemical_list: D001426:Bacterial Proteins; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease