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Spectroscopy
Volume 17 (2003), Issue 2-3, Pages 377-398
http://dx.doi.org/10.1155/2003/801452

Using UV-Vis. Linear Dichroism to Study the Orientation of Molecular Probes and Biomolecules in Lipidic Membranes

M. A. R. B. Castanho,1,2 S. Lopes,2 and M. Fernandes3

1Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Campo Grande C8, 1749-016 Lisboa, Portugal
2Centro de Química Física Molecular, Complexo I, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
3Departamento de Química Física, Universidad de Murcia, 30071 Murcia, Spain

Copyright © 2003 Hindawi Publishing Corporation. 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

Linear dichroism methodologies are based on the different interaction of molecules with linearly polarized light depending on their orientation. Theoretical predictions are used to conclude on the orientation of selected molecules relative to their neighbours (usually an organized matrix). In the specific case of linear dichroism methodologies applied to data obtained from UV-Vis. spectroscopic techniques, the orientational distribution function of the chromophores' electronic transition moment can be calculated and converted to the molecular axis distribution function. In this paper, the orientation of molecular probes and biomolecules in model systems of biomembranes (lipidic matrixes) is explored and illustrated using common membrane probes (trans-parinaric acid, a cyanine and laurdan) and polyene antibiotics (Amphotericin B and Nystatin). The emphasis is on the technique and methodologies themselves, rather than the scientific impact of the attained distributions. The main addressed items are: (1) How to adapt a common UV-Vis. spectrophotometer and spectrofluorimeter to perform linear dicrhoism experiments?, (2) Sample preparation, and (3) Data analysis (including artefacts corrections).