Halarnkar P P, Nelson J H, Heisler C R, Blomquist G J
Arch Biochem Biophys. 1985 Feb 1;236(2):526-34. doi: 10.1016/0003-9861(85)90655-1.
Carbon-13 NMR and radiotracer studies were used to determine the precursor to methylmalonate and to study the metabolism of propionate in the cockroach Periplaneta americana. [3,4,5-13C3]Valine labeled carbons 3, 4, and 26 of 3-methylpentacosane, indicating that valine was metabolized via propionyl-CoA to methylmalonyl-CoA and served as the methyl branch unit precursor. Potassium [2-13C]propionate labeled the odd-numbered carbons of hydrocarbons and potassium [3-13C]propionate labeled the even-numbered carbons of hydrocarbons in this insect. This labeling pattern indicates that propionate is metabolized to acetate, with carbon-2 of propionate becoming the methyl carbon of acetate and carbon-3 of propionate becoming the carboxyl carbon of acetate. In vivo studies in which products were separated by HPLC showed that [2-14C]propionate was readily metabolized to acetate. The radioactivity from sodium [1-14C]propionate was not incorporated into succinate nor into any other tricarboxylic acid cycle intermediate, indicating that propionate was not metabolized via methylmalonate to succinate. Similarly, [1-14C]propionate did not label acetate. An experiment designed to determine the subcellular localization of the enzymes involved in converting propionate to acetate showed that they were located in the mitochondrial fraction. Data from both in vivo and in vitro studies as a function of time indicated that propionate was converted directly to acetate and did not first go through tricarboxylic acid cycle intermediates. These data demonstrate a novel pathway of propionate metabolism in insects.
利用碳-13核磁共振和放射性示踪剂研究来确定丙二酸单酰的前体,并研究蜚蠊美洲大蠊中丙酸的代谢。[3,4,5-13C3]缬氨酸标记了3-甲基二十五烷的碳3、碳4和碳26,表明缬氨酸通过丙酰辅酶A代谢为甲基丙二酰辅酶A,并作为甲基支链单位前体。[2-13C]丙酸钾标记了该昆虫中烃类的奇数碳,[3-13C]丙酸钾标记了烃类的偶数碳。这种标记模式表明,丙酸代谢为乙酸,丙酸的碳2成为乙酸的甲基碳,丙酸的碳3成为乙酸的羧基碳。通过高效液相色谱法分离产物的体内研究表明,[2-14C]丙酸很容易代谢为乙酸。[1-14C]丙酸钠的放射性既没有掺入琥珀酸中,也没有掺入任何其他三羧酸循环中间产物中,这表明丙酸不是通过丙二酸单酰代谢为琥珀酸。同样,[1-14C]丙酸也没有标记乙酸。一项旨在确定将丙酸转化为乙酸所涉及酶的亚细胞定位的实验表明,这些酶位于线粒体部分。体内和体外研究的数据随时间变化表明,丙酸直接转化为乙酸,并不首先经过三羧酸循环中间产物。这些数据证明了昆虫中丙酸代谢的一条新途径。