Dynamic nuclear polarization-enhanced solid-state NMR of a 13C-labeled signal peptide bound to lipid-reconstituted sec translocon

Reggie, Lenica; Lopez, Jakob J.; Collinson, Ian; Glaubitz, Clemens; Lorch, Mark

Abstract

Dynamic nuclear polarization (DNP) has made it possible to record 2D double-quantum-filtered (DQF) solid-state NMR (ssNMR) spectra of a signal peptide bound to a lipid-reconstituted SecYEG translocon complex. The small quantity of peptide in the sample (∼40 nmol) normally prohibits multidimensional ssNMR experiments. Such small amounts are not the exception, because for samples involving membrane proteins, most of the limited sample space is occupied by lipids. As a consequence, a conventional 2D DQF ssNMR spectrum with the sample used here would require many weeks if not months of measurement time. With the help of DNP, however, we were able to acquire such a 2D spectrum within 20 h. This development opens up new possibilities for membrane protein studies, particularly in the exploitation of high-resolution spectroscopy and the assignment of individual amino acid signals, in this case for a signal peptide bound to the translocon complex.

Citation

Reggie, L., Lopez, J. J., Collinson, I., Glaubitz, C., & Lorch, M. (2011). Dynamic nuclear polarization-enhanced solid-state NMR of a 13C-labeled signal peptide bound to lipid-reconstituted sec translocon. Journal of the American Chemical Society, 133(47), 19084-19086. https://doi.org/10.1021/ja209378h

• Journal Article Type: Article
• Acceptance Date: Feb 1, 2011
• Online Publication Date: Nov 1, 2011
• Publication Date: Nov 30, 2011
• Deposit Date: Nov 13, 2014
• Journal: Journal Of The American Chemical Society
• Print ISSN: 0002-7863
• Electronic ISSN: 1520-5126
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 133
• Issue: 47
• Pages: 19084-19086
• DOI: https://doi.org/10.1021/ja209378h
• Keywords: Colloid and Surface Chemistry; Biochemistry; General Chemistry; Catalysis
• Public URL: https://hull-repository.worktribe.com/output/466770
• Publisher URL: https://doi.org/10.1021/ja209378h