Advanced search
Start date
Betweenand

Development and use of fluorescence techniques for the study of protein conformational changes

Grant number: 14/00661-0
Support type:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): August 11, 2014
Effective date (End): July 20, 2015
Field of knowledge:Biological Sciences - Biophysics - Molecular Biophysics
Principal researcher:Vitor Marcelo Silveira Bueno Brandão de Oliveira
Grantee:Marcelo Ferreira Marcondes Machado
Supervisor abroad: Jaap Broos
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Research place: University of Groningen, Netherlands  
Associated to the scholarship:11/20941-9 - DEVELOPMENT AND USE OF FLUORESCENCE TECHNIQUES FOR THE STUDY OF PROTEIN CONFORMATIONAL CHANGES, BP.PD

Abstract

The main goal of the present project is to study the hinge movement that the peptidase, timet oligopeptidase (TOP) may undergo upon substrate or inhibitor binding. Fluorescence techniques will be used to study this conformational change in great detail. TOP constructs will be prepared equipped with two donor-donor fluorescent probes (Trp residues) or donor-acceptor probes (5-F Trp - p-nitro-phenylalanine) pairs, one at each side of the central deep channel of the TOP structure. Fluorescence experiments, in steady state and time-resolved mode will inform us about the hinge movement in this enzyme and the impact of different substrates and inhibitors on it. The kinetics of the hinge movement will be investigated using the same construct via stopped-flow fluorescence spectroscopy. A better understanding of the hinge movement will allow us to tune the activity of this enzyme and provides a good basis for designing new efficient inhibitors. We foresee that once we successful demonstrate the potential of this approach especially the 5-FTrp - p-nitro-phenylalanine donor-acceptor pair as outlined in this application, this approach will be adopted by other researchers aiming to study protein conformational changes. (AU)

News published in Agência FAPESP Newsletter about the scholarship:
Articles published in other media outlets (0 total):
More itemsLess items
VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)

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)
OLIVEIRA-SOUZA, WELLINGTON P.; BRONZE, FELLIPE; BROOS, JAAP; MARCONDES, MARCELO F. M.; OLIVEIRA, VITOR. On the efficient bio-incorporation of 5-hydroxy-tryptophan in recombinant proteins expressed in Escherichia coli with T7 RNA polymerase-based vectors. Biochemical and Biophysical Research Communications, v. 492, n. 3, p. 343-348, OCT 21 2017. Web of Science Citations: 1.
MARCONDES, M. F. M.; ALVES, F. M.; ASSIS, D. M.; HIRATA, I. Y.; JULIANO, L.; OLIVEIRA, V.; JULIANO, M. A. Substrate specificity of mitochondrial intermediate peptidase analysed by a support-bound peptide library. FEBS OPEN BIO, v. 5, p. 429-436, 2015. Web of Science Citations: 1.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.