Time-resolved fluorescence investigations of the interaction of the voltage-sensitive probe RH421 with lipid membranes and proteins

Biochemistry
N V VisserR J Clarke

Abstract

Fluorescence lifetimes and fluorescence anisotropy decays of the voltage-sensitive styryl-pyridinium dye RH421 have been measured in the presence of dimyristoylphosphatidylcholine vesicles, the water soluble enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco), and Na+,K(+)-ATPase-containing membrane fragments. The effect of an intramembrane electric field on the photophysical properties of the dye was investigated by the binding of the hydrophobic ion tetraphenylborate (TPB) to the membrane. TPB was found to significantly increase the average fluorescence lifetime and the order of membrane-bound dye. The increase in fluorescence lifetime is consistent with reorientation of the dye further into the membrane interior. The increase in order may be attributed to an electric field-induced alignment of the dye molecules. From the values of the rotational diffusion constant experimentally determined, the expected response time of the dye to an applied electric field can be calculated for a reorientational mechanism to be on the order of tens of nanoseconds. Experiments with rubisco showed that the dye interacts strongly with the protein. In this case, the dye is so tightly bound that it has almost no independent motion an...Continue Reading

Citations

Jan 10, 2009·European Biophysics Journal : EBJ·Thomas Starke-Peterkovic, Ronald J Clarke
Apr 28, 2004·Bioelectrochemistry·Hans-Jürgen Apell
Dec 24, 1997·Annals of the New York Academy of Sciences·I KlodosF Cornelius
Oct 8, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Hirak ChakrabortyAmitabha Chattopadhyay
Dec 21, 2006·Biochimica Et Biophysica Acta·Gaëlle Le GoffRonald J Clarke
Sep 18, 2014·Biophysical Journal·Laura J MaresRonald J Clarke
Jan 31, 1997·Biochimica Et Biophysica Acta·R J Clarke, D J Kane
Jun 29, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Maria MatsonBengt Nordén
Aug 23, 2003·AJR. American Journal of Roentgenology·Leonard Berlin
Apr 22, 2008·The Journal of Physical Chemistry. B·T H Nguyen Pham, Ronald J Clarke
Feb 19, 2004·The Journal of Physical Chemistry. B·Marlon J HinnerPeter Fromherz
Feb 24, 2001·Advances in Colloid and Interface Science·R J Clarke
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May 16, 2021·Biochimica Et Biophysica Acta. Biomembranes·Anna ChmielińskaAgnieszka Polit
Dec 6, 2012·The Journal of Physical Chemistry Letters·Mehmet OzbilRajeev Prabhakar
Dec 15, 2010·Biochemistry·Hans-Jürgen ApellDaniel Sauerbrunn
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