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Real-time measurement of H/D exchange in a microdialysis Raman quartz cell

✍ Scribed by Pedro Carmona; Marina Molina

Publisher
John Wiley and Sons
Year
2009
Tongue
English
Weight
184 KB
Volume
40
Category
Article
ISSN
0377-0486

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✦ Synopsis

Abstract

We present here a novel quartz cell for monitoring H/D exchange in biomolecules using Raman spectroscopy. This cell is combined with a syringe to pump heavy water through a hollow microdialysis fibre, which is inserted into the cell. The deuterium efflux into the sample has been studied as a function of the molecular weight cut‐off of the microdialysis fibre and compared with other microcell systems comprising conventional glass capillaries. The fastest D~2~O efflux that we have obtained (k~d~ = 0.38 Β± 0.008 min^βˆ’1^) permits to measure exchange rates of 2.5 min^βˆ’1^ or less. Application of this cell to deuterium exchange in glyceraldehyde‐3‐phosphate dehydrogenase reveals a class of H‐atoms highly resistant to deuteration, which is consistent with a previous infrared study on this protein. Copyright Β© 2009 John Wiley & Sons, Ltd.

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