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Spectroscopy: An International Journal
Volume 27, Article ID 791342, 6 pages
http://dx.doi.org/10.1155/2012/791342

A New Method for D 2 O / H 2 O Exchange in Infrared Spectroscopy of Proteins

1Laboratorio di Biochimica e Biofisica, Dipartimento di Biologia, Università di Bologna, 40126 Bologna, Italy
2Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia, c/o Dipartimento di Fisica, Università di Bologna, 40127 Bologna, Italy
3LASIR, UMR CNRS 8516, Bat C5, Université Lille 1, 59655 Villeneuve d'Ascq, France
4Service de Bioénergétique Biologie Structurale et Mécanismes, URA CNRS 2096, IBITeC-S, Bat 532, CEA-Saclay, 91191 Gif-sur-Yvette Cedex, France

Copyright © 2012 Marco Malferrari et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

In this paper, we describe a new method to obtain D2O/H2O exchange in photosynthetic reaction centres from Rhodobacter sphaeroides. The method is characterized by: (i) a very high efficiency of the isotopic replacement; (ii) an extremely low amount of D2O needed; (iii) the short time required for dehydration and D2O rehydration; (iv) the possibility of controlling concomitantly the hydration state of the sample. The proposed method can be applied to other proteins.