Premium
Real‐time measurement of H/D exchange in a microdialysis Raman quartz cell
Author(s) -
Carmona Pedro,
Molina Marina
Publication year - 2009
Publication title -
journal of raman spectroscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.748
H-Index - 110
eISSN - 1097-4555
pISSN - 0377-0486
DOI - 10.1002/jrs.2283
Subject(s) - raman spectroscopy , chemistry , microdialysis , analytical chemistry (journal) , deuterium , chromatography , extracellular , biochemistry , physics , quantum mechanics , optics
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 ± 0.008 min −1 ) permits to measure exchange rates of 2.5 min −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 © 2009 John Wiley & Sons, Ltd.