title: Probing slowly exchanging protein systems via ¹³Cα-CEST: monitoring folding of the Im7 protein.
abstract: A¹³C(α) chemical exchange saturation transfer based experiment is presented for the study of protein systems undergoing slow interconversion between an 'observable' ground state and one or more 'invisible' excited states. Here a labeling strategy whereby [2-(13)C]-glucose is the sole carbon source is exploited, producing proteins with ¹³C at the C(α) position, while the majority of residues remain unlabeled at CO or C(β). The new experiment is demonstrated with an application to the folding reaction of the Im7 protein that involves an on-pathway excited state. The obtained excited state (13)C(α) chemical shifts are cross validated by comparison to values extracted from analysis of CPMG relaxation dispersion profiles, establishing the utility of the methodology.
authors: Department of Molecular Genetics, The University of Toronto, Toronto, ON, M5S 1A8, Canada.
authors: Alexandar L
authors: AL
authors: Hansen
authors: Guillaume
authors: G
authors: Bouvignies
authors: Lewis E
authors: LE
authors: Kay
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry*; D003198:Computer Simulation; D029968:Escherichia coli Proteins / Q000737:chemistry*; D009010:Monte Carlo Method; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017510:Protein Folding
chemical_list: D002247:Carbon Isotopes; D002352:Carrier Proteins; C509253:E colicin-binding immunity protein Im7, E coli; D029968:Escherichia coli Proteins
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