Monitoring methanol-induced protein unfolding by fluorescence anisotropy measurements of covalently labelled rhodamine probe

Antonin Soleilhac (1) , Franck Bertorelle (1) , Philippe Dugourd (1) , Marion Girod (2) , Rodolphe Antoine (1)

1 ILM - Institut Lumière Matière [Villeurbanne]
2 ISA-ANABIO-MS - Analyse biomoléculaire par spectrométrie de masse - Biological Analysis by Mass Spectrometry

We describe the use of an extrinsic fluorophore (Rhodamine B isothiocyanate) as a versatile probe to measure rotational motions of proteins. To illustrate the usefulness of this probe, we describe the fluorescence anisotropy values of this fluorophore covalently linked to myoglobin protein measured in aqueous solutions of increased methanol content. Methanol-induced unfolding is revealed by the transition from constrained to free rotation of the covalently attached rhodamine B fluorophore. Graphical abstract Constrained to Free a) b) MGLSDGEWQQVLNVWGKVEA DIAGHGQEVLIRLFTGHPET LEKFDKFKHLKTEAEMKASE DLKKHGTVVLTALGGILKKK GHHEAELKPLAQSHATKHKI PIKYLEFISDAIIHVLHSKH PGDFGADAQGAMTKALELFR NDIAAKYKELGFQG 2 Introduction:

Domaines: Chimie analytique

Antonin Soleilhac, Franck Bertorelle, Philippe Dugourd, Marion Girod, Rodolphe Antoine. Monitoring methanol-induced protein unfolding by fluorescence anisotropy measurements of covalently labelled rhodamine probe. The European Physical Journal D : Atomic, molecular, optical and plasma physics, 2017, 71 (6), ⟨10.1140/epjd/e2017-70760-3⟩. ⟨hal-01569441⟩