Tang Wai-Kwan, Chan Chi-Bun, Cheng Christopher H K, Fong Wing-Ping
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, NT, Hong Kong, China.
FEBS Lett. 2005 Jul 4;579(17):3759-64. doi: 10.1016/j.febslet.2005.05.070.
Subsequent to our earlier report on the first purification of antiquitin protein from seabream liver and demonstration of its enzymatic activity [FEBS Letters 516 (2002) 183-186], we report herein the cloning of its full-length cDNA sequence. The open reading frame encodes a protein of 511 amino acids. Results of RT-PCR indicate that antiquitin is highly expressed in both the seabream liver and kidney. Transfection studies in cultured eukaryotic cells provided further evidence that it is a cytosolic protein. Bacterial expression of the enzyme was also performed. The purified recombinant protein was demonstrated to exhibit similar kinetic properties as the native enzyme.
继我们之前关于首次从鲷鱼肝脏中纯化抗泛素蛋白并证明其酶活性的报告[《欧洲生物化学学会联合会快报》516 (2002) 183 - 186]之后，我们在此报告其全长cDNA序列的克隆。开放阅读框编码一个由511个氨基酸组成的蛋白质。逆转录聚合酶链反应结果表明抗泛素在鲷鱼的肝脏和肾脏中均高度表达。在培养的真核细胞中进行的转染研究进一步证明它是一种胞质蛋白。还进行了该酶的细菌表达。纯化的重组蛋白被证明具有与天然酶相似的动力学特性。
