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Structural Alterations of Human Serum Albumin Caused by Glycative and Oxidative Stressors Revealed by Circular Dichroism Analysis
Department of Internal Medicine, University of Genova, Genova 16132, Italy
Institute for Macromolecular Studies-ISMAC, National Research Council-CNR, Genova 16149, Italy
Department of Experimental Medicine, Section of General Pathology, University of Genova, Genova 16132, Italy
This author deceased on 25 August 2010.
* Author to whom correspondence should be addressed.
Received: 11 March 2013; in revised form: 22 April 2013 / Accepted: 2 May 2013 / Published: 23 May 2013
Abstract: The aim of this work was to evaluate the ability of oxidative and glycative stressors to modify properties of human serum albumin (HSA) by analyzing markers of glycation (pentosidine) and oxidation (advanced oxidative protein products (AOPPs)) and assessing fluorescence and circular dichroism. HSA was incubated for up to 21 days with ribose, ascorbic acid (AA) and diethylenetriamine pentacetate (DTPA) in various combinations in order to evaluate influences of these substances on the structure of HSA. Ribose was included as a strong glycative molecule, AA as a modulator of oxidative stress, and DTPA as an inhibitor of metal-catalyzed oxidation. Ribose induced a significant increase in pentosidine levels. AA and DTPA prevented the accumulation of pentosidine, especially at later time points. Ribose induced a mild increase in AOPP formation, while AA was a strong inducer of AOPP formation. Ribose, in combination with AA, further increased the formation of AOPP. DTPA prevented the AA-induced generation of AOPP. Ribose was also a potent inducer of fluorescence at 335nm ex/385nm em, which is typical of pentosidine. AA and DTPA prevented this fluorescence. Circular dichroism showed complex results, in which AA and DTPA were strong modifiers of the percentages of the alpha-helical structure of HSA, while ribose affected the structure of HSA only at later time points.
Keywords: ribose; ascorbic acid; pentosidine; fluorescence; circular dichroism
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Monacelli, F.; Storace, D.; D'Arrigo, C.; Sanguineti, R.; Borghi, R.; Pacini, D.; Furfaro, A.L.; Pronzato, M.A.; Odetti, P.; Traverso, N. Structural Alterations of Human Serum Albumin Caused by Glycative and Oxidative Stressors Revealed by Circular Dichroism Analysis. Int. J. Mol. Sci. 2013, 14, 10694-10709.
Monacelli F, Storace D, D'Arrigo C, Sanguineti R, Borghi R, Pacini D, Furfaro AL, Pronzato MA, Odetti P, Traverso N. Structural Alterations of Human Serum Albumin Caused by Glycative and Oxidative Stressors Revealed by Circular Dichroism Analysis. International Journal of Molecular Sciences. 2013; 14(6):10694-10709.
Monacelli, Fiammetta; Storace, Daniela; D'Arrigo, Cristina; Sanguineti, Roberta; Borghi, Roberta; Pacini, Davide; Furfaro, Anna L.; Pronzato, Maria A.; Odetti, Patrizio; Traverso, Nicola. 2013. "Structural Alterations of Human Serum Albumin Caused by Glycative and Oxidative Stressors Revealed by Circular Dichroism Analysis." Int. J. Mol. Sci. 14, no. 6: 10694-10709.