Characterizing Protein Hydration Dynamics Using Solution NMR Spectroscopy

Overview

Journal Methods Enzymol

Specialty Biochemistry

Date 2019 Jan 15

PMID 30638541

Citations 7

Authors

Christine Jorge

Bryan S Marques

Kathleen G Valentine

A Joshua Wand

Affiliations

Soon will be listed here.

Abstract

Protein hydration is a critical aspect of protein stability, folding, and function and yet remains difficult to characterize experimentally. Solution NMR offers a route to a site-resolved view of the dynamics of protein-water interactions through the nuclear Overhauser effects between hydration water and the protein in the laboratory (NOE) and rotating (ROE) frames of reference. However, several artifacts and limitations including contaminating contributions from bulk water potentially plague this general approach and the corruption of measured NOEs and ROEs by hydrogen exchange-relayed magnetization. Fortunately, encapsulation of single protein molecules within the water core of a reverse micelle overcomes these limitations. The main advantages are the suppression hydrogen exchange and elimination of bulk water. Here we detail guidelines for the preparation solutions of encapsulated proteins that are suitable for characterization by NOE and ROE spectroscopy. Emphasis is placed on understanding the contribution of detected NOE intensity arising from magnetization relayed by hydrogen exchange. Various aspects of fitting obtained NOE, selectively decoupled NOE, and ROE time courses are illustrated.

Citing Articles

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Protein conformational entropy is not slaved to water.

Marques B, Stetz M, Jorge C, Valentine K, Wand A, Nucci N Sci Rep. 2020; 10(1):17587.

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