Partial purification and functional reconstitution of GDP sensitive brown adipose tissue mitochondria uncoupling protein using octyl glucoside.
A partial purification of the uncoupling protein of brown adipose tissue mitochondria (BATM) was achieved by extraction of BATM with 40 mM octyl glucoside, followed by affinity chromatography on ATP-agarose. The isolated protein was functionally reconstituted into liposomes using octyl glucoside dialysis. Proteoliposomes containing the uncoupling protein had an increased proton or chloride conductance when subjected to a valinomycin-induced potassium diffusion potential. The increased ion conductance was consistently found to be inhibited by 200 microM GDP.
Strieleman
P J
PJ
Schalinske
K L
KL
Shrago
E
E
D002001:Adipose Tissue, Brown / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins; D002712:Chlorides / Q000378:metabolism; D002846:Chromatography, Affinity; D006224:Cricetinae; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005960:Glucosides; D006153:Guanosine Diphosphate / Q000378:metabolism; D007473:Ion Channels; D008081:Liposomes / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins; D011522:Protons; D000071256:Uncoupling Protein 1
D002352:Carrier Proteins; D002712:Chlorides; D003902:Detergents; D005960:Glucosides; D007473:Ion Channels; D008081:Liposomes; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D011522:Protons; D000071256:Uncoupling Protein 1; D006153:Guanosine Diphosphate; C018619:octyl-beta-D-glucoside