pH-temperature interactions on protein function and hibernation: GDP binding to brown adipose tissue mitochondria.
1. [3H]GDP binding to the uncoupling protein of brown adipose tissue was determined on mitochondria isolated from hibernating European hamsters, at two temperatures, 35 and 15 degrees C, and four values of 25pH (pH corrected to 25 degrees C): 6.4, 6.8, 7.2 and 7.6, encompassing the physiological range of pH. Buffer composition was adjusted to get the same pH-temperature relationship as for mammalian blood, in which this relationship is mainly determined by protein imidazole buffers. 2. The maximal binding capacity was independent both of temperature and pH. The dissociation constant KD was highly pH-dependent, but was independent of temperature when 25pH was held constant. Under these conditions, the uncoupling protein thus fully conserves its regulatory properties over the temperature range studied (eurythermal adaptation). 3. The temperature coefficient of the apparent pK' for the pH effect (-0.012 +/- 0.004) differed significantly from that of GDP terminal phosphoryl group, but not from that of blood protein imidazole buffer groups, in good agreement with the imidazole alphastat theory. 4. The results indicate that GDP reaction with the protein involves an electrostatic binding with a titratable group of the protein, probably a sulfhydryl. 5. pH modulation of the uncoupling of brown adipose tissue mitochondria probably permits a reversible control of thermogenesis in the hibernation cycle, heat dissipation being inhibited by respiratory acidosis in deep hibernation, but facilitated by the hyperventilation of arousal.
Malan
A
A
CNRS, Laboratoire d'Etude des Régulations Physiologiques, Strasbourg, France.
Mioskowski
E
E
D002001:Adipose Tissue, Brown / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D006224:Cricetinae; D006150:Guanine Nucleotides / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006605:Hibernation; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D011506:Proteins / Q000502:physiology*; D013696:Temperature
D006150:Guanine Nucleotides; D011506:Proteins; D006153:Guanosine Diphosphate; D006859:Hydrogen