Summary
Amide-resolved, hydrogen-deuterium exchange from bee venom melittin reconstituted in fully hydrated vesicles suspended in D2O buffer was measured using a technique involving (1) trapping samples throughout an exchange time course by rapid freezing and lyophilization; and (2) dissolving the dried peptide/lipid mixtures in deuteromethanol to record high-resolution spectra using semiselective excitation pulses to select peptide amide signals in the presence of large excess lipid signals. Two-dimensional, amide-selective GaussNOESY and fingerprint-selective off-diagonal PingCOSY spectra are shown to be suitable for rapid acquisition of amide-selective spectra, obtained throughout a time course of amide exchange in the membrane-bound state. Membrane-reconstituted melittin is shown to contain two sequences of exchange-stable amides, corresponding to helical regions on either side of the single proline residue.
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Dempsey, C.E. Amide-resolved hydrogen—deuterium exchange measurements from membrane-reconstituted polypeptides using exchange trapping and semiselective two-dimensional NMR. J Biomol NMR 4, 879–884 (1994). https://doi.org/10.1007/BF00398417
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DOI: https://doi.org/10.1007/BF00398417