Lysine 92 amino acid residue of USP46, a gene associated with 'behavioral despair' in mice, influences the deubiquitinating enzyme activity.
Lysine 92 amino acid residue of USP46, a gene associated with 'behavioral despair' in mice, influences the deubiquitinating enzyme activity.
复制标题
DOI：
10.1371/journal.pone.0026297
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Wang MW
中科院分区：
文献类型：
--
作者：
Zhang W;Tian QB;Li QK;Wang JM;Wang CN;Liu T;Liu DW;Wang MW
去泛素化酶（DUBs）通过从特定的蛋白质底物上切割泛素来调节多种细胞功能。泛素特异性蛋白酶46（Ubiquitin-Specific Protease 46，USP 46）是近年来发现的一个数量性状基因，在小鼠悬尾试验和强迫游泳试验中起着重要的作用。USP 46中赖氨酸密码子（Lys 92）缺失的小鼠在不可避免的应激下表现出“行为绝望”的丧失，以及昼夜行为节律和GABA能系统的异常。然而，这种缺失是否影响酶活性尚不清楚。在这里，我们表明，USP 46具有去泛素化酶活性检测USP裂解试验使用GST-Ub 52作为模型底物。有趣的是，与野生型相比，Lys 92缺失突变体导致去泛素化酶活性降低27.04%。我们还使用实时RT-PCR测定了Usp 46在大鼠组织中的相对表达水平。Usp 46 mRNA在包括脑在内的各种组织中均有表达，其中在脾脏中的表达最高。此外，与大鼠USP 46一样，人和小鼠USP 46对模型底物都有活性，表明USP裂解试验是检测USP 46去泛素化酶活性的简单方法。这些结果表明，USP 46的Lys 92缺失可能影响酶活性，从而提供了酶如何调节精神疾病发病机制的分子线索。
Deubiquitinating enzymes (DUBs) regulate diverse cellular functions by their activity of cleaving ubiquitin from specific protein substrates. Ubiquitin-Specific Protease 46 (USP46) has recently been identified as a quantitative trait gene responsible for immobility in the tail suspension test and forced swimming test in mice. Mice with a lysine codon (Lys 92) deletion in USP46 exhibited loss of ‘behavioral despair’ under inescapable stresses in addition to abnormalities in circadian behavioral rhythms and the GABAergic system. However, whether this deletion affects enzyme activity is unknown. Here we show that USP46 has deubiquitinating enzyme activity detected by USP cleavage assay using GST-Ub52 as a model substrate. Interestingly, compared to wild type, the Lys 92 deletion mutant resulted in a decreased deubiquitinating enzyme activity of 27.04%. We also determined the relative expression levels of Usp46 in rat tissues using real-time RT-PCR. Usp46 mRNA was expressed in various tissues examined including brain, with the highest expression in spleen. In addition, like rat USP46, both human and mouse USP46 are active toward to the model substrate, indicating the USP cleavage assay is a simple method for testing the deubiquitinating enzyme activity of USP46. These results suggest that the Lys 92 deletion of USP46 could influence enzyme activity and thereby provide a molecular clue how the enzyme regulating the pathogenesis of mental illnesses.
登录
查看更多内容
影响因子：
62.1
作者：
Reyes-Turcu, Francisca E.;Wilkinson, Keith D.
通讯作者：
Wilkinson, Keith D.
影响因子：
3.9
作者：
Li, Yonghong;Schrodi, Steven;Grupe, Andrew
通讯作者：
Grupe, Andrew
影响因子：
4.8
作者：
Cohn, Martin A.;Kee, Younghoon;D'Andrea, Alan D.
通讯作者：
D'Andrea, Alan D.
影响因子：
8
作者：
Bernardi, R;Liebermann, DA;Hoffman, B
通讯作者：
Hoffman, B
影响因子：
4.7
作者：
Anderson, C;Crimmins, S;Wilson, SM
通讯作者：
Wilson, SM