Molecular recognition phenomena play crucial roles in a number of biological processes. Thus the elucidation of these processes deserves the understanding of the basic forces that determine the involved protein-protein or small molecule-protein interactions. This aim determines a growing need of methodologies for studying any specific biomolecular event as well as the structural elements that drive the process. In order to determine the equilibrium and/or kinetic constants for binding, these techniques must factor the concentrations of free probe and target molecules, as well as of the corresponding complex. Many methodologies are well suited to get information on the complex formation as well as on the funzionalizing high structures. However a multimethodological approach is often essential to give a deeper insight in the understanding of the recognition process, taking advantage of the peculiarity of each methodology. Here we report the use of biochromatography, circular dichroism, and optical biosensors in key activities in drug discovery and developments, e.g. identification of protein function modulators, and early determination of ADME parameters. In particular, applications of the different methodologies will be presented for determining the distribution parameters of anticancer and antiviral drugs, studying the drug binding to target proteins, and getting information on the association of functional proteins into homo and hetero-oligomers (1-4). As far as the distribution parameters determination is concerned, the binding of drugs to albumins from different species have been characterized in terms of bound fraction, affinity constant, binding area localization, and stereochemistry of the drug-protein complex. The relevance of target discovery and validation will be then addressed by applying optical biosensor technique and circular dichroism spectroscopy to anticancer drugs. Finally, the characterization of the molecular interactions between herpes simplex virus (HSV) cellular receptors and glycoproteins showing receptor-binding activity will be reported. The use of different methodologies resulted of great relevance for understanding the glycoprotein-mediated interference to infection and then to get information on the mechanism involved in the virus entry. 1. S. Cimitan, M.T. Lindgren, C. Bertucci, U.H. Danielson J.Med.Chem., 2005, 48:7282-9. 2. T. Gianni, A. Piccoli, C. Bertucci, G. Campadelli-Fiume. J Virol., 2006, 80:2216-24. 3. G.A. Ascoli, E. Domenici, C. Bertucci, Chirality, 2006, 18:667-79. 4. B. Sanavio, A. Piccoli, T. Gianni, C. Bertucci, BBA-PRO, 2007, in press.

BIORECOGNITION STUDIES: MULTIMETHODOLOGICAL APPROACH

BERTUCCI, CARLO;PICCOLI, ANGELA;PISTOLOZZI, MARCO
2007

Abstract

Molecular recognition phenomena play crucial roles in a number of biological processes. Thus the elucidation of these processes deserves the understanding of the basic forces that determine the involved protein-protein or small molecule-protein interactions. This aim determines a growing need of methodologies for studying any specific biomolecular event as well as the structural elements that drive the process. In order to determine the equilibrium and/or kinetic constants for binding, these techniques must factor the concentrations of free probe and target molecules, as well as of the corresponding complex. Many methodologies are well suited to get information on the complex formation as well as on the funzionalizing high structures. However a multimethodological approach is often essential to give a deeper insight in the understanding of the recognition process, taking advantage of the peculiarity of each methodology. Here we report the use of biochromatography, circular dichroism, and optical biosensors in key activities in drug discovery and developments, e.g. identification of protein function modulators, and early determination of ADME parameters. In particular, applications of the different methodologies will be presented for determining the distribution parameters of anticancer and antiviral drugs, studying the drug binding to target proteins, and getting information on the association of functional proteins into homo and hetero-oligomers (1-4). As far as the distribution parameters determination is concerned, the binding of drugs to albumins from different species have been characterized in terms of bound fraction, affinity constant, binding area localization, and stereochemistry of the drug-protein complex. The relevance of target discovery and validation will be then addressed by applying optical biosensor technique and circular dichroism spectroscopy to anticancer drugs. Finally, the characterization of the molecular interactions between herpes simplex virus (HSV) cellular receptors and glycoproteins showing receptor-binding activity will be reported. The use of different methodologies resulted of great relevance for understanding the glycoprotein-mediated interference to infection and then to get information on the mechanism involved in the virus entry. 1. S. Cimitan, M.T. Lindgren, C. Bertucci, U.H. Danielson J.Med.Chem., 2005, 48:7282-9. 2. T. Gianni, A. Piccoli, C. Bertucci, G. Campadelli-Fiume. J Virol., 2006, 80:2216-24. 3. G.A. Ascoli, E. Domenici, C. Bertucci, Chirality, 2006, 18:667-79. 4. B. Sanavio, A. Piccoli, T. Gianni, C. Bertucci, BBA-PRO, 2007, in press.
12th International Meeting on Recent Developments in Pharmaceutical Analysis RDPA 2007
18
18
Bertucci C.; Piccoli A.; Pistolozzi M.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/54192
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