Fiche publication
Date publication
février 2017
Journal
The journal of physical chemistry. B
Auteurs
Membres identifiés du Cancéropôle Est :
Pr MELY Yves
,
Mr HUMBERT Nicolas
,
Pr DIDIER Pascal
Tous les auteurs :
Ghose A, Maltsev OV, Humbert N, Hintermann L, Arntz Y, Naumov P, Mély Y, Didier P
Lien Pubmed
Résumé
In this work, we used firefly oxyluciferin (OxyLH2) and its polarity-dependent fluorescence mechanism as a sensitive tool to monitor biomolecular interactions. The chromophores, OxyLH2, and its two analogues, 4-MeOxyLH and 4,6'-DMeOxyL, were modified trough carboxylic functionalization and then coupled to the N-terminus part of Tat and NCp7 peptides of human immunodeficiency virus type-1 (HIV-1). The photophysical properties of the labeled peptides were studied in live cells as well as in complex with different oligonucleotides in solution. By monitoring the emission properties of these derivatives we were able, for the first time, to study in vitro biomolecular interactions using oxyluciferin as a sensor. As an additional application, cyclopropyl-oxyluciferin (5,5-Cpr-OxyLH) was site-specifically conjugated to the thiol group (Cys-232) of the human protein α-1 antytripsin to investigate its interaction with porcine pancreatic elastase. Our data demonstrate that OxyLH2 and its derivatives can be used as fluorescence reporters for monitoring biomolecular interactions.
Mots clés
Animals, Fireflies, Fluorescent Dyes, chemical synthesis, HIV-1, HeLa Cells, Humans, Hydrogen-Ion Concentration, Indoles, chemical synthesis, Oligodeoxyribonucleotides, chemistry, Pancreatic Elastase, chemistry, Protein Binding, Pyrazines, chemical synthesis, Swine, alpha 1-Antitrypsin, chemistry, gag Gene Products, Human Immunodeficiency Virus, chemistry, tat Gene Products, Human Immunodeficiency Virus, chemistry
Référence
J Phys Chem B. 2017 Feb;121(7):1566-1575