Removal of bound Triton X-100 from purified bovine heart cytochrome bc1.
Cytochrome bc(1) 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 bc(1) 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 bc(1) 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 bc(1) retained nearly full activity, absorption spectra, subunit, and phospholipid composition.
Varhac
Rastislav
R
Department of Biochemistry, University of Texas Health Science Center, San Antonio, TX 78229, USA.
Robinson
Neal C
NC
Musatov
Andrej
A
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003902:Detergents / Q000737:chemistry*; D014450:Electron Transport Complex III / Q000737:chemistry / Q000302:isolation & purification*; D008929:Mitochondria, Heart / Q000201:enzymology; D017830:Octoxynol / Q000737:chemistry*; D020358:Sodium Cholate / Q000737:chemistry
D003902:Detergents; D017830:Octoxynol; D014450:Electron Transport Complex III; D020358:Sodium Cholate