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• W2009821000 abstract "Atom-resolved real-time studies of kinetic processes in proteins have been hampered in the past by the lack of experimental techniques that yield sufficient temporal and atomic resolution. Here we present band-selective optimized flip-angle short transient (SOFAST) real-time 2D NMR spectroscopy, a method that allows simultaneous observation of reaction kinetics for a large number of nuclear sites along the polypeptide chain of a protein with an unprecedented time resolution of a few seconds. SOFAST real-time 2D NMR spectroscopy combines fast NMR data acquisition techniques with rapid sample mixing inside the NMR magnet to initiate the kinetic event. We demonstrate the use of SOFAST real-time 2D NMR to monitor the conformational transition of alpha-lactalbumin from a molten globular to the native state for a large number of amide sites along the polypeptide chain. The kinetic behavior observed for the disappearance of the molten globule and the appearance of the native state is monoexponential and uniform along the polypeptide chain. This observation confirms previous findings that a single transition state ensemble controls folding of alpha-lactalbumin from the molten globule to the native state. In a second application, the spontaneous unfolding of native ubiquitin under nondenaturing conditions is characterized by amide hydrogen exchange rate constants measured at high pH by using SOFAST real-time 2D NMR. Our data reveal that ubiquitin unfolds in a gradual manner with distinct unfolding regimes." @default.
• W2009821000 created "2016-06-24" @default.
• W2009821000 creator A5003272694 @default.
• W2009821000 creator A5069093700 @default.
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• W2009821000 date "2007-07-03" @default.
• W2009821000 modified "2023-10-16" @default.
• W2009821000 title "Protein folding and unfolding studied at atomic resolution by fast two-dimensional NMR spectroscopy" @default.
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• W2009821000 doi "https://doi.org/10.1073/pnas.0702069104" @default.
• W2009821000 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2040886" @default.
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