Entendendo as interações moleculares e a estabilidade da proteína verde fluorescente (GFP) em líquidos iônicos sob condições de estresse

• Grant number: 18/20833-0
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Start date: January 29, 2019
• End date: May 28, 2019
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Jorge Pereira
• Grantee: Carolina Falaschi Saponi
• Supervisor: Tamar Greaves
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Institution abroad: RMIT University, Melbourne, Australia
• Associated to the scholarship: 18/01858-2 - Study of the stabilizing effect of different solvents on the green fluorescent protein structure produced by Escherichia coli, BP.IC

Abstract

The Green Fluorescent Protein (GFP) has intense and natural fluorescence, being easily quantifiable and having an excellent application as biosensor and biomarker. However, the manifestation of its fluorescence is dependent of the maintenance of the integrity of its protein structure, which is very sensitive to different conditions, such as high temperatures, extreme pH or presence of denaturing substances. The instability of GFP under various conditions, allied with its high cost of production, restricts its use to small-scale applications. Thus, the study of solvents that protect the structure of GFP under stress conditions is essential to allow its use on a large scale. The objective of this project is to study the interactions that occur at a molecular level between GFP and aqueous solutions of different solvents and conditions in order to reveal the interactions between proteins and ILs. In particular ionic liquids (ILs) at different concentrations, temperatures, pH and in the presence of denaturing substances will be explored. This study has the major goal of improving and broadening the applications of GFP in the medical and industrial fields. The fluorescence intensity will be evaluated in association with in-depth studies of its secondary and tertiary structure, using spectroscopy and light scattering techniques.

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)

DOS SANTOS, NATHALIA VIEIRA; SAPONI, CAROLINA FALASCHI; RYAN, TIMOTHY M.; PRIMO, FERNANDO L.; GREAVES, TAMAR L.; PEREIRA, JORGE F. B.. Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors. International Journal of Biological Macromolecules, v. 164, p. 3474-3484, DEC 1 2020. (18/20833-0, 14/16424-7, 14/19793-3, 18/06576-5, 16/07529-5, 18/50009-8, 18/01858-2)

V. VERISSIMO, NATHALIA; SAPONI, CAROLINA F.; RYAN, TIMOTHY M.; GREAVES, TAMAR L.; PEREIRA, JORGE F. B.. Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity. GREEN CHEMICAL ENGINEERING, v. 2, n. 4, p. 11-pg., 2021-12-01. (18/06576-5, 18/25511-1, 14/16424-7, 20/14144-8, 18/50009-8, 14/19793-3, 18/01858-2, 18/20833-0, 16/07529-5)