How can deuterium replace hydrogen?

Hydrogen–deuterium exchange (also called H–D or H/D exchange) is a chemical reaction in which a covalently bonded hydrogen atom is replaced by a deuterium atom, or vice versa.

How do you do deuterium exchange?

Due to the acidic nature of α hydrogens they can be exchanged with deuterium by reaction with D2O (heavy water). The process is accelerated by the addition of an acid or base; an excess of D2O is required. The end result is the complete exchange of all α hydrogens with deuterium.

How many hydrogen atoms will deuterium atoms exchange?

In the second option the alpha-carbon is the one attached to the first carbon of benzene and it contains two alpha-hydrogens hence, a total of two alpha hydrogens will be exchanged with deuterium as: Hence, maximum number of hydrogen atoms exchanged with deuterium is 5.

How is HDX data Analysed?

The most commonly used strategy for HDX-MS is to digest the proteins into peptides and analyze them using mass spectrometry. This ensures complete sequence coverage and captures region-specific information from the protein.

What happens when hydrogen is exchanged for deuterium?

Hydrogen–Deuterium Exchange. H/D exchange experiments are based on the chemical reaction of replacing covalently bonded hydrogens with deuterium atoms to reveal the tertiary structure information of proteins.

What is hydrogen deuterium exchange mass spectrometry ( HDX-MS )?

Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful technique that can provide insights into protein behavior by serving as a link between structure, conformational dynamics and function 1.

How is deuterium exchange used to study protein structure?

Hydrogen–deuterium exchange methods have proven to be a powerful addition to other biophysical techniques available to study protein structure and dynamics.

How is the rate of deuterium incorporation measured?

The rate of deuterium incorporation is characteristic of local structure and dynamics. H/D exchange coupled with mass spectrometry measures the deuteration rate. H/D exchange experiments with different reaction periods are quenched by lowering pH to ~2.5. Quenching is followed by proteolysis, chromatographic separation, and mass analysis.