Hiltunen J K, Kauppinen R A, Nuutinen E M, Peuhkurinen K J, Hassinen I E
Biochem J. 1980 Jun 15;188(3):725-9. doi: 10.1042/bj1880725.
The metabolism of four short-chain odd-number-carbon fatty acids, pentanoate, pent-4-enoate, propionate and acrylate, was studied in isolated rat heart mitochondria incubated in [14C]bicarbonate buffer. Under these conditions pentanoate was metabolized with a concomitant accumulation of malate and incorporation of 14CO2 into non-volatile compounds. The metabolism of propionate to tricarboxylic acid-cycle intermediates required the addition of ATP and oligomycin. After addition of a small amount of rotenone to the incubation medium, pent-4-enoate was metabolized with an increase in malate from less than 3 nmol/mg of protein to 34.0 +/- 1.5 nmol/mg in 40 min, during which time the amount of 14CO2 fixed in acid-stable compounds increased from 1.56 +/- 0.30 to 41.1 +/- 2.6 nmol/mg of protein. Acrylate was not metabolized under any of the conditions tested. The results show that cardiac mitochondria must have an enzyme system that is capable of reducing the double bond of either pent-4-enoate or its metabolities. That the metabolism of pent-4-enoate occurs through a reductive step and energy-dependent carboxylation is evident from the requirement for NAD+ reduction by partial inhibition of the mitochondrial respiratory chain and the presence of ATP and CO2. The results do not enable us to say whether the compound reduced is pent-4-enoyl-CoA or acryloyl-CoA.
在含有[¹⁴C]碳酸氢盐缓冲液的离体大鼠心脏线粒体中,研究了四种短链奇数碳脂肪酸(戊酸、4-戊烯酸、丙酸和丙烯酸)的代谢情况。在这些条件下,戊酸代谢时伴随着苹果酸的积累以及¹⁴CO₂掺入非挥发性化合物中。丙酸代谢为三羧酸循环中间产物需要添加ATP和寡霉素。在孵育介质中加入少量鱼藤酮后,4-戊烯酸开始代谢,40分钟内苹果酸含量从低于3 nmol/mg蛋白质增加到34.0±1.5 nmol/mg,在此期间,酸稳定化合物中固定的¹⁴CO₂量从1.56±0.30增加到41.1±2.6 nmol/mg蛋白质。在任何测试条件下,丙烯酸都不发生代谢。结果表明,心脏线粒体必定具有一种能够还原4-戊烯酸或其代谢产物双键的酶系统。通过部分抑制线粒体呼吸链对NAD⁺还原的需求以及ATP和CO₂的存在,可知4-戊烯酸的代谢是通过还原步骤和能量依赖性羧化作用进行的。结果无法让我们确定被还原的化合物是4-戊烯酰辅酶A还是丙烯酰辅酶A。