TY - JOUR
T1 - Continuous fluorescence-based measurement of redox-driven sodium ion translocation.
AU - Muras, Valentin
AU - Claussen , Björn
AU - Karuppasamy, Manikandan
AU - Steuber, Julia
AU - Schaffitzel, Christiane
PY - 2014/8/15
Y1 - 2014/8/15
N2 - Investigation of the mechanism of sodium ion pumping enzymes requires methods to follow the translocation of sodium ions by the purified and reconstituted proteins in vitro. Here, we describe a protocol that allows following the accumulation of Na(+) in proteoliposomes by the Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) from Vibrio cholerae using the sodium-sensitive fluorophor sodium green. In the presence of a regenerative system for its substrate NADH, the Na(+)-NQR accumulates Na(+) in the proteoliposomes which is visible as a change in fluorescence.
AB - Investigation of the mechanism of sodium ion pumping enzymes requires methods to follow the translocation of sodium ions by the purified and reconstituted proteins in vitro. Here, we describe a protocol that allows following the accumulation of Na(+) in proteoliposomes by the Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) from Vibrio cholerae using the sodium-sensitive fluorophor sodium green. In the presence of a regenerative system for its substrate NADH, the Na(+)-NQR accumulates Na(+) in the proteoliposomes which is visible as a change in fluorescence.
U2 - 10.1016/j.ab.2014.05.012
DO - 10.1016/j.ab.2014.05.012
M3 - Article (Academic Journal)
C2 - 24862438
SN - 0003-2697
VL - 459
SP - 53
EP - 55
JO - Analytical Biochemistry
JF - Analytical Biochemistry
ER -