Brooks S P, Storey K B
Department of Biology, Carleton University, Ottawa, Ontario, Canada.
Am J Physiol. 1993 Dec;265(6 Pt 2):R1380-6. doi: 10.1152/ajpregu.1993.265.6.R1380.
Changes in de novo protein synthesis and protein phosphorylation were monitored during anoxia and recovery in the red-eared slider Trachemys (= Pseudemys) scripta elegans. Time courses of 35S-radiolabeled methionine incorporation into acid-precipitable material showed an increase up to 5 h postinjection and remained constant after this time. Comparison of the total and acid-precipitable 35S label incorporation into tissues from 20-h control, anoxic, and recovering animals showed differences between these groups: total radioactivity in brain was 2.9-fold lower in recovering turtles, whereas protein-associated radioactivity was 2.4-fold higher in anoxic liver, 2.3-fold lower in recovering skeletal muscle, and 3.7-fold lower in recovering brain tissue. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radiolabeled proteins showed the existence of a newly synthesized protein band (relative molecular mass = 72 kDa) that was apparent only in 20-h recovering liver and skeletal muscle. Use of 32P labeling to monitor changes in protein phosphorylation patterns during anoxia revealed 1.6-, 1.4-, and 1.5-fold increases in 32P incorporation in anoxic brain, heart, and liver, respectively. Changes in protein phosphorylation were localized to the plasma membrane and cytosolic fractions in brain and to the cytosolic fraction in liver.
在红耳龟（滑龟属=伪龟属）秀丽锦龟的缺氧及恢复过程中，监测了从头合成蛋白质和蛋白质磷酸化的变化。35S放射性标记的甲硫氨酸掺入酸沉淀物质的时间进程显示，注射后5小时内有所增加，此后保持恒定。对20小时对照、缺氧和恢复中的动物组织中总35S标记和酸沉淀35S标记掺入情况的比较显示，这些组之间存在差异：恢复中的海龟大脑中的总放射性降低了2.9倍，而缺氧肝脏中与蛋白质相关的放射性增加了2.4倍，恢复中的骨骼肌中降低了2.3倍，恢复中的脑组织中降低了3.7倍。放射性标记蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示存在一条新合成的蛋白带（相对分子质量=72 kDa），仅在恢复20小时的肝脏和骨骼肌中明显。使用32P标记监测缺氧期间蛋白质磷酸化模式的变化，结果显示缺氧的大脑、心脏和肝脏中32P掺入分别增加了1.6倍、1.4倍和1.5倍。蛋白质磷酸化的变化定位于大脑中的质膜和胞质部分以及肝脏中的胞质部分。
