Release and fixation of CO2 by guinea-pig kidney tubules metabolizing aspartate.
1. The metabolism of L-[U-14C]aspartate, L-[1-14C]aspartate and L-[4-14C]aspartate was studied in isolated guinea-pig kidney tubules. 2. Oxidation of C-1 plus that of C-4 of aspartate accounted for 90-92% of the CO2 released from aspartate, whereas oxidation of the inner carbon atoms of aspartate (which occurs beyond the 2-oxoglutarate dehydrogenase step) represented only 8-10% of aspartate carbon oxidation. 3. The formation of [1-14C]glutamine and [1-14C]glutamate from [1-14C]aspartate and [4-14C]aspartate indicated that about one-third of the oxaloacetate synthesized from aspartate underwent randomization at the level of fumarate. 4. With [U-14C]aspartate as substrate, the percentage of the C-1 of glutamate and glutamine found radiolabelled after 60 min of incubation was 92.7% and 47.5% in the absence and the presence of bicarbonate respectively. 5. That CO2 fixation occurred at high rates in the presence of bicarbonate was demonstrated by incubating tubules with aspartate plus [14C]bicarbonate; under this condition, the label fixed was found in C-1 of glutamate, glutamine and aspartate, as well as in C-4 of aspartate, demonstrating not only randomization of aspartate carbon but also aspartate resynthesis secondary to oxaloacetate cycling via phosphoenolpyruvate carboxykinase, pyruvate kinase and pyruvate carboxylase. 6. The importance of CO2 fixation in glutamine synthesis from aspartate is discussed in relation to the possible role of the guinea-pig kidney in systemic acid-base regulation in vivo.
Martin
G
G
Laboratoire de Physiologie Rénale et Métabolique, CNRS URA 1177, Faculté de Médecine Alexis Carrel, Lyon, France.
Michoudet
C
C
Vincent
N
N
Baverel
G
G
D000086:Acetazolamide / Q000494:pharmacology; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism*; D001639:Bicarbonates / Q000378:metabolism; D002021:Buffers; D002245:Carbon Dioxide / Q000378:metabolism*; D002250:Carbon Radioisotopes; D005973:Glutamine / Q000096:biosynthesis; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007684:Kidney Tubules / Q000187:drug effects / Q000378:metabolism*; D010084:Oxidation-Reduction; D011865:Radioisotope Dilution Technique; D012964:Sodium / Q000378:metabolism; D017693:Sodium Bicarbonate
D001639:Bicarbonates; D002021:Buffers; D002250:Carbon Radioisotopes; D005973:Glutamine; D002245:Carbon Dioxide; D001224:Aspartic Acid; D017693:Sodium Bicarbonate; D012964:Sodium; D000086:Acetazolamide