Issue 74, 2020

Tm filtering by 1H-methyl labeling in a deuterated protein for pulsed double electron–electron resonance EPR

Abstract

Modulating the phase-memory relaxation time (Tm) of a spin label by introducing 1H-methyl groups in a perdeuterated protein background is used in DEER experiments to assign interactions in multimodal P(r) distributions. Proof of principle is demonstrated using Protein A where one nitroxide label occupies two distinct regions of conformational space. The presence of a single protonated methyl group in close proximity (4–8 Å) to only one of the two nitroxide rotamer ensembles results in a selective and substantial decrease in Tm, manifested by differential decay of the peak intensities in the bimodal P(r) distance distribution as a function of the total dipolar evolution time. This form of Tm filtering will facilitate DEER structural analysis of biomolecular systems with three spin labels, including complexes involving multimeric proteins.

Graphical abstract: T m filtering by 1H-methyl labeling in a deuterated protein for pulsed double electron–electron resonance EPR

Supplementary files

Article information

Article type
Communication
Submitted
23 iyn 2020
Accepted
12 avq 2020
First published
12 avq 2020

Chem. Commun., 2020,56, 10890-10893

T m filtering by 1H-methyl labeling in a deuterated protein for pulsed double electron–electron resonance EPR

T. Schmidt and G. M. Clore, Chem. Commun., 2020, 56, 10890 DOI: 10.1039/D0CC04369A

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements