TOAC spin-labeled peptides tailored for DNP-NMR studies in lipid membrane environments

S Zhu; E Kachooei; JR Harmer; LJ Brown; F Separovic; MA Sani

Journal article

TOAC spin-labeled peptides tailored for DNP-NMR studies in lipid membrane environments

S Zhu, E Kachooei, JR Harmer, LJ Brown, F Separovic, MA Sani

Biophysical Journal | Published : 2021

DOI: 10.1016/j.bpj.2021.08.040

Abstract

The benefit of combining in-cell solid-state dynamic nuclear polarization (DNP) NMR and cryogenic temperatures is providing sufficient signal/noise and preservation of bacterial integrity via cryoprotection to enable in situ biophysical studies of antimicrobial peptides. The radical source required for DNP was delivered into cells by adding a nitroxide-tagged peptide based on the antimicrobial peptide maculatin 1.1 (Mac1). In this study, the structure, localization, and signal enhancement properties of a single (T-MacW) and double (T-T-MacW) TOAC (2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid) spin-labeled Mac1 analogs were determined within micelles or lipid vesicles. The s..

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University of Melbourne Researchers

Frances Separovic Author

Marco Sani Author

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Grants

Awarded by Australian Research Council

Funding Acknowledgements

[ "The authors gratefully acknowledge Dr. John Karas (University of Melbourne) for his help with the peptide synthesis. S. Z. thanks the Albert Shimmins Fund for Postgraduate Writing-up Award.", "This research was funded by the Australian Research Council Discovery Project grant DP160100959 and DP190101506 to F.S. and Linkage Infrastructure, Equipment and Facilities grant LE160100120 to F.S. and M.-A.S. DNP-NMR was accessed at the Bio21 Institute NMR facility." ]