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Deubiquitinating enzyme amino acid profiling reveals a class of ubiquitin esterases [Biochemistry]
Proceedings of the National Academy of Sciences of the United States of America ( IF 11.1 ) Pub Date : 2021-01-26 , DOI: 10.1073/pnas.2006947118
Virginia De Cesare 1 , Daniel Carbajo Lopez 1 , Peter D Mabbitt 1 , Adam J Fletcher 1 , Mathieu Soetens 1 , Odetta Antico 1 , Nicola T Wood 1 , Satpal Virdee 2
Affiliation  

The reversibility of ubiquitination by the action of deubiquitinating enzymes (DUBs) serves as an important regulatory layer within the ubiquitin system. Approximately 100 DUBs are encoded by the human genome, and many have been implicated with pathologies, including neurodegeneration and cancer. Non-lysine ubiquitination is chemically distinct, and its physiological importance is emerging. Here, we couple chemically and chemoenzymatically synthesized ubiquitinated lysine and threonine model substrates to a mass spectrometry-based DUB assay. Using this platform, we profile two-thirds of known catalytically active DUBs for threonine esterase and lysine isopeptidase activity and find that most DUBs demonstrate dual selectivity. However, with two anomalous exceptions, the ovarian tumor domain DUB class demonstrates specific (iso)peptidase activity. Strikingly, we find the Machado–Joseph disease (MJD) class to be unappreciated non-lysine DUBs with highly specific ubiquitin esterase activity rivaling the efficiency of the most active isopeptidases. Esterase activity is dependent on the canonical catalytic triad, but proximal hydrophobic residues appear to be general determinants of non-lysine activity. Our findings also suggest that ubiquitin esters have appreciable cellular stability and that non-lysine ubiquitination is an integral component of the ubiquitin system. Its regulatory sophistication is likely to rival that of canonical ubiquitination.



中文翻译:

去泛素化酶氨基酸谱揭示了一类泛素酯酶 [生物化学]

通过去泛素化酶 (DUB) 的作用实现的泛素化可逆性是泛素系统中的一个重要调节层。大约 100 个 DUB 由人类基因组编码,其中许多与病理有关,包括神经变性和癌症。非赖氨酸泛素化在化学上是不同的,其生理重要性正在显现。在这里,我们将化学和化学酶法合成的泛素化赖氨酸和苏氨酸模型底物与基于质谱的 DUB 测定相结合。使用该平台,我们分析了三分之二的已知催化活性 DUB 的苏氨酸酯酶和赖氨酸异肽酶活性,并发现大多数 DUB 表现出双重选择性。然而,除了两个异常例外,卵巢肿瘤结构域 DUB 类表现出特异性(异)肽酶活性。引人注目的是,我们发现 Machado-Joseph 病 (MJD) 类是未被重视的非赖氨酸 DUB,具有高度特异性的泛素酯酶活性,可与最活跃的异肽酶的效率相媲美。酯酶活性取决于典型的催化三联体,但近端疏水残基似乎是非赖氨酸活性的一般决定因素。我们的研究结果还表明,泛素酯具有明显的细胞稳定性,并且非赖氨酸泛素化是泛素系统的一个组成部分。它的监管复杂性很可能与典型的泛素化相媲美。酯酶活性取决于典型的催化三联体,但近端疏水残基似乎是非赖氨酸活性的一般决定因素。我们的研究结果还表明,泛素酯具有明显的细胞稳定性,并且非赖氨酸泛素化是泛素系统的一个组成部分。它的监管复杂性很可能与典型的泛素化相媲美。酯酶活性取决于典型的催化三联体,但近端疏水残基似乎是非赖氨酸活性的一般决定因素。我们的研究结果还表明,泛素酯具有明显的细胞稳定性,并且非赖氨酸泛素化是泛素系统的一个组成部分。它的监管复杂性很可能与典型的泛素化相媲美。

更新日期:2021-01-22
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