A study of GDP binding to purified thermogenin protein from brown adipose tissue.
1. The binding of GDP to purified thermogenin protein was studied by using fluorescence spectroscopy and equilibrium dialysis. 2. GDP binding to thermogenin diminished fluorescence emission in a concentration-dependent manner that exhibited saturation. 3. Kd values for binding of nucleoside di- and tri-phosphates were lower than those for nucleoside monophosphates. 4. The GDP-induced fluorescence quenching was decreased by increasing pH, but the apparent Kd was unaltered by pH changes. 5. Equilibrium dialysis showed a Kd change from 3 to 6 microM when the pH was increased from 6.6 to 8.5. 6. The apparent pK of the fluorescence changes induced by pH (8.3) was identical with the apparent pK of the GDP-binding response. 7. The data are consistent with the existence of protonated and non-protonated forms of thermogenin protein that both bind GDP.
D002001:Adipose Tissue, Brown / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins; D009711:Nucleotides / Q000378:metabolism; D051381:Rats; D011924:Rats, Zucker; D013050:Spectrometry, Fluorescence; D000071256:Uncoupling Protein 1
D002352:Carrier Proteins; D006150:Guanine Nucleotides; D007473:Ion Channels; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D009711:Nucleotides; D000071256:Uncoupling Protein 1; D006153:Guanosine Diphosphate; D019204:GTP-Binding Proteins