A Note on the use of Steady-State Fluorescence Quenching to Quantify Nanoparticle-Protein Interactions

Autor(es)

Sousa, Alioscka A.

• Tipo de documento: Artigo Científico
• Fonte: Journal of Fluorescence; v. 25, n. 6, p. 1567-1575, NOV 2015.
• Citações Web of Science: 7

Resumo

Steady-state fluorescence quenching is a commonly used technique to investigate the interactions between proteins and nanoparticles, providing quantitative information on binding affinity, stoichiometry and cooperativity. However, a failure to account for the limitations and pitfalls of the methodology can lead to significant errors in data analysis and interpretation. Thus, in this communication we first draw attention to a few common pitfalls in the use of fluorescence quenching to study nanoparticle-protein interactions. For example, we discuss a frequent mistake in the use of the Hill equation to determine cooperativity. We also test using both simulated and experimental data the application of a model-independent method of analysis to generate true thermodynamic nanoparticle-protein binding isotherms. This model-free approach allows for a quantitative description of the interactions independent of assumptions about the nature of the binding process {[}Bujalowski W, Lohman TM (1987) Biochemistry 26: 3099; Schwarz G (2000) Biophys. Chem. 86: 119]. (AU)

• Processo FAPESP: 13/18481-5 - Interações biológicas de uma biblioteca de nanopartículas de ouro com aplicações na nanomedicina
• Beneficiário: Alioscka Augusto Sousa
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores