Holmes R P
Department of Urology Bowman, Gray School of Medicine, Winston-Salem, NC 27157.
Biochim Biophys Acta. 1993 Aug 20;1158(1):47-51. doi: 10.1016/0304-4165(93)90095-p.
There have been several reports over the past decade of the presence of the glyoxylate cycle enzymes, isocitrate lyase and malate synthase, in animal tissues. Reaction products in these assays have been measured principally by chromatographic separation of isotopes or by colorimetric procedures. In this report more sensitive and accurate HPLC and HPCE analyses were used to detect enzymatic activity. Reversed phase HPLC revealed the absence of detectable isocitrate lyase activity in guinea pig, rat and chick embryonic liver. The formation of several other alpha-keto acids was detected and this may account for the previously reported activities. Using HPCE to monitor malate formation malate synthase activity was not detected in these tissues. These results indicate that when assaying enzyme activities in crude tissue homogenates specific methods for the identification of end products are required.
在过去十年里,有几份报告指出动物组织中存在乙醛酸循环酶,即异柠檬酸裂解酶和苹果酸合酶。这些测定中的反应产物主要通过同位素的色谱分离或比色法来测量。在本报告中,使用了更灵敏和准确的高效液相色谱(HPLC)和高效毛细管电泳(HPCE)分析来检测酶活性。反相高效液相色谱显示,豚鼠、大鼠和鸡胚胎肝脏中未检测到可检测到的异柠檬酸裂解酶活性。检测到了其他几种α-酮酸的形成,这可能解释了先前报道的活性。使用高效毛细管电泳监测苹果酸的形成,在这些组织中未检测到苹果酸合酶活性。这些结果表明,在粗组织匀浆中测定酶活性时,需要特定的终产物鉴定方法。