The use of click chemistry as a tool for studying protein modifications by cholesterol aldehydes

• Grant number: 12/16145-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: January 15, 2013
• End date: January 14, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Sayuri Miyamoto
• Grantee: Thiago Cardoso Genaro de Mattos
• Supervisor: Ned A. Porter
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Vanderbilt University (VU), United States

Abstract

Cholesterol is a neutral lipid widely found in the cell membrane, where it influences in the membrane fluidity, organization and in cell signaling events. Since cholesterol is known to play structural and regulatory functions in the membrane, its physiological concentration is strictly controlled by the cell. An increase in the mitochondrial cholesterol, for instance, has been associated with a depletion in the mitochondrial glutathione levels, which is known to decrease the cell antioxidant defenses. This scenario will increase the oxidative stress and, consequently, enhance the cholesterol oxidation. The oxidation of cholesterol can generate reactive aldehydes. Several studies reported elevated levels of cholesterol aldehydes in atherosclerotic plaques and in brain tissue of patients bearing neurodegenerative diseases. Moreover, evidences suggest the ability of these aldehydes to covalently react with proteins, leading to conformational changes and malfunction. On the other hand, the extension of protein modifications by cholesterol aldehydes and their implications to the cell are poorly known. In this sense, the development of tools and techniques with the purpose of identifying modified proteins is of great interest, since it can help understanding their roles in cell signaling events and in the development of several diseases. Therefore, this project aims to study protein modifications promoted by cholesterol aldehydes using click chemistry. To achieve these goals, we intend to synthetize modified cholesterol aldehydes, characterize their reactions with peptides and proteins and identify their major targets inside the cell. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

WINDSOR, KATHERINE; GENARO-MATTOS, THIAGO C.; MIYAMOTO, SAYURI; STEC, DONALD F.; KIM, HYE-YOUNG H.; TALLMAN, KERI A.; PORTER, NED A.. Assay of Protein and Peptide Adducts of Cholesterol Ozonolysis Products by Hydrophobic and Click Enrichment Methods. Chemical Research in Toxicology, v. 27, n. 10, p. 1757-1768, OCT 2014. (12/16145-5)