Fedorchuk Tat'yana, Rudenko Natalia, Ignatova Lyudmila, Ivanov Boris
Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino 142290, Russia.
Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino 142290, Russia.
J Plant Physiol. 2014 Jul 1;171(11):903-6. doi: 10.1016/j.jplph.2014.02.009. Epub 2014 Mar 11.
Supernatant obtained after high-speed centrifugation of disrupted thylakoids that had been washed free from extrathylakoid carbonic anhydrases demonstrated carbonic anhydrase activity that was inhibited by the specific inhibitors acetazolamide and ethoxyzolamide. A distinctive feature of the effect of Triton X-100 on this activity also suggested that the source of the activity is a soluble protein. Native electrophoresis of a preparation obtained using chromatography with agarose/mafenide as an affinity sorbent revealed one protein band with carbonic anhydrase activity. The same protein was revealed in a mutant deficient in soluble stromal carbonic anhydrase β-CA1, and this indicated that the newly revealed carbonic anhydrase is not a product of the At3g01500 gene. These data imply the presence of soluble carbonic anhydrase in the thylakoid lumen of higher plants.
从已洗去类囊体膜外碳酸酐酶的破碎类囊体高速离心后获得的上清液，表现出被特异性抑制剂乙酰唑胺和乙氧唑胺抑制的碳酸酐酶活性。Triton X-100对该活性的影响的一个显著特征也表明该活性的来源是一种可溶性蛋白质。使用琼脂糖/甲磺灭脓作为亲和吸附剂进行色谱法获得的制剂的天然电泳显示出一条具有碳酸酐酶活性的蛋白带。在缺乏可溶性基质碳酸酐酶β-CA1的突变体中也发现了相同的蛋白质，这表明新发现的碳酸酐酶不是At3g01500基因的产物。这些数据意味着高等植物类囊体腔中存在可溶性碳酸酐酶。
