TY - JOUR
T1 - Removal of bound Triton X-100 from purified bovine heart cytochrome bc1
AU - Varhač, Rastislav
AU - Robinson, Neal C.
AU - Musatov, Andrej
PY - 2009/12/15
Y1 - 2009/12/15
N2 - Cytochrome bc1 isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc1 is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc1 retained nearly full activity, absorption spectra, subunit, and phospholipid composition.
AB - Cytochrome bc1 isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc1 is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc1 retained nearly full activity, absorption spectra, subunit, and phospholipid composition.
UR - http://www.scopus.com/inward/record.url?scp=70349785067&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70349785067&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2009.08.042
DO - 10.1016/j.ab.2009.08.042
M3 - Article
C2 - 19733142
AN - SCOPUS:70349785067
VL - 395
SP - 268
EP - 270
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 2
ER -