Hock B
Institut für Biologie der Universität Tübingen, Tübingen, Deutschland.
Planta. 1970 Mar;93(1):26-38. doi: 10.1007/BF00387649.
Previously, it was deduced from inhibitor experiments that isocitrate lyase (EC 4.1.3.1.) is synthesized de novo in watermelon cotyledons during the first 3 days of germination, which explains the sharp increase of activity during this period. The following decrease of activity was interpreted as the result of a limited half life of the enzyme molecule (Hock and Beevers, 1966).This hypothesis has been confirmed now by density labeling experiments of isocitrate lyase with deuterium. Seedlings grown from day 0 on D2O (80 vol. %) contained a heavier enzyme at the time of maximum activity than control seedlings grown on H2O (Fig. 6). No incorporation of deuterium into isocitrate lyase, however, was detectable when the cotyledons were labeled only from day 3 1/2 on, i.e. after the stage of maximum activity had been passed (Fig. 10), in spite of the fact that D2O was taken up from the cotyledons in considerable quantities. -These results prove at the same time that density labeling of the isocitrate lyase during early stages of germination was a result of de novo synthesis rather than a mere artifact produced by isotopic exchange.An improved method for the purification of isocitrate lyase from higher plants is introduced.
此前,通过抑制剂实验推断,异柠檬酸裂解酶(EC 4.1.3.1.)在西瓜子叶萌发的前3天中是重新合成的,这解释了该时期酶活性的急剧增加。随后酶活性的下降被解释为酶分子半衰期有限的结果(霍克和比弗斯,1966年)。现在,通过用氘对异柠檬酸裂解酶进行密度标记实验,这一假设得到了证实。在D2O(80体积%)中从第0天开始生长的幼苗,在酶活性达到最大值时,其所含的酶比在H2O中生长的对照幼苗的酶更重(图6)。然而,当仅从第3.5天开始对子叶进行标记时,即酶活性达到最大值的阶段过后(图10),尽管子叶大量吸收了D2O,但未检测到氘掺入异柠檬酸裂解酶中。这些结果同时证明,在萌发早期对异柠檬酸裂解酶进行密度标记是重新合成的结果,而不是由同位素交换产生的假象。本文介绍了一种从高等植物中纯化异柠檬酸裂解酶的改进方法。