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ChemBioChem

Volumn 10, Issue 13, 2009, Pages 2177-2181

In situ monitoring of backbone thioester exchange by 19F NMR

(4) Pomerantz, William C a Hadley, Erik B a Fry, Charles G a Gellman, Samuel H a

a UNIVERSITY OF WISCONSIN MADISON (United States)

Author keywords

Backbone thioester exchange; Cooled coil; Fluorine; NMR spectroscopy; Protein folding

Indexed keywords

DEPSIPEPTIDE; POLYPEPTIDE; THIOESTER; THIOL;

ANALYTIC METHOD; ARTICLE; CONTROLLED STUDY; EQUILIBRIUM CONSTANT; FLUORINE NUCLEAR MAGNETIC RESONANCE; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; PH; PRIORITY JOURNAL; PROTEIN CONFORMATION; PROTEIN FOLDING; PROTEIN MOTIF; PROTEIN STABILITY; SCREENING; SENSITIVITY AND SPECIFICITY;

AMINO ACID SEQUENCE; ESTERS; FLUORINE; MAGNETIC RESONANCE SPECTROSCOPY; MOLECULAR SEQUENCE DATA; PEPTIDES; PROTEIN STRUCTURE, TERTIARY; SULFHYDRYL COMPOUNDS;

EID: 70349387405 PISSN: 14394227 EISSN: 14397633 Source Type: Journal
DOI: 10.1002/cbic.200900380 Document Type: Article

Times cited : (6)

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- See the Supporting Information.

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- The HPLC-based BTE equilibrium analyses were conducted as described previously (refs. 3 and 5, each thiol and thioester contained a Tyr residue, and KBTE was measured by determining the relative populations of the four components by using HPLC after equilibration was complete. The 19F NMR-based BTE equilibrium analyses were conducted with fluorine-labeled components; KBTE was measured by determining the relative populations of the four components by using integration of 19F resonances after equilibration was complete. In contrast to this latter approach, Δδ 19F(f2Phe)-based ΔGFold estimates are obtained simply by comparing d 19F(f2Phe) for the full-length thiodepsipeptide and the C-terminal peptide thiol. It is not necessary that equilibrium be attained for this estimation
- 2Phe) for the full-length thiodepsipeptide and the C-terminal peptide thiol. It is not necessary that equilibrium be attained for this estimation.

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- HPLC was used to measure Δ. Details may be found in the Supporting Information
- 19F NMR-based estimation method did indeed correctly identify the most and least stable coiled coils within this set. Details may be found in the Supporting Information.
- 19F NMR-based estimation method did indeed correctly identify the most and least stable coiled coils within this set

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