Joint determination by Brownian dynamics and fluorescence quenching of the in-depth location profile of biomolecules in membranes.
The in-depth molar distribution function of fluorophores is revealed by a new methodology for fluorescence quenching data analysis in membranes. Brownian dynamics simulation was used to study the in-depth location profile of quenchers. A Lorentzian profile was reached. Since the Stern-Volmer equation is valid at every depth in the membrane for low quencher concentrations, the molar distribution of the fluorophore (also regarded as a Lorentzian) can be achieved. The average location and the broadness of the fluorophore distribution can be calculated. The importance of the knowledge of the location width is demonstrated and discussed, since this parameter reveals important conclusions on structural features of the interaction of membranes with probes and biomolecules (e.g., conformational freedom in proteins), as well as photophysical properties (e.g., differential fluorophore quantum yields). Subsequent use of this methodology by the reader does not, necessarily, involve the performance of simulations and is not limited to the use of Lorentzian function distributions.
Fernandes
M X
MX
Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Lisbon, Portugal.
García de la Torre
J
J
Castanho
M A R B
MA
D002462:Cell Membrane; D005453:Fluorescence; D005456:Fluorescent Dyes; D007700:Kinetics; D008051:Lipid Bilayers; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry; D008433:Mathematics; D008565:Membrane Proteins / Q000737:chemistry*; D008961:Models, Structural; D008962:Models, Theoretical; D010713:Phosphatidylcholines; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008055:Lipids; D008081:Liposomes; D008565:Membrane Proteins; D010713:Phosphatidylcholines