Hydrogen Deuterium Exchange

From LEAP

	Hydrogen Deuterium Exchange
	Application Type
	Sample Prep and Inject
	Application ID
	HD-x PAL
	Description
	Scheduling and Sample Prep for HD-x

Overview

H/D-X PAL™ is an easy-to-use, system that provides an automated process for the scheduling and experimental execution of H/D-X experimental workflow. By use of the advanced LEAP Shell scheduling software experimental design is simplified and reliable. Synchronous reagent addition and sample labeling steps are automatically scheduled to increase throughput and produce high quality data.

Protein structural studies

Information on protein structure can be determined by one of 3 commonly used techniques:

1. Crystalography

2. NMR

3. HD Exchange

In cases where there is insufficient sample (NMR) or the protein will not crystalize, Hydrogen Deuterium exchange is the only option to obtain structural data. It relies on the accurate measurement of the degree of labeling of a protein by deuterated hydrogen during a precisely measured labeling interval. The deuterium is exchanged with hydrogen atoms on the exposed amine backbone of the protein and depending on the which regions are exposed and how the protein is folded, differences are seen in the MS spectra of the digested protein when compared with non-labelled proteins.

Labelling time is important (exposure time of the protein to Deuterium) as is prevention of "back-exchange" after the labelling has occured. This is the natural tendancy of normal hydrogen to replace the deuterium after the label is removed. This is minimized by reducing the temperature of the mixture to close to zero degress Celsius.