Identification of key features required for efficient S-acylation and plasma membrane targeting of sprouty-2.
Identification of key features required for efficient S-acylation and plasma membrane targeting of sprouty-2.
复制标题
鉴定sprougty-2的有效S-酰化和质膜靶向所需的关键特征。
DOI：
10.1242/jcs.249664
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发表时间：
2020-11-05
影响因子：
4
通讯作者：
Chamberlain LH
中科院分区：
文献类型：
--
作者：
Locatelli C;Lemonidis K;Salaun C;Tomkinson NCO;Chamberlain LH
Sprouty-2是一种重要的生长因子信号调节因子和肿瘤抑制蛋白。该蛋白的特征是一个富含半胱氨酸的结构域，含有26个半胱氨酸残基，被S酰化修饰。在本研究中，我们发现Sprouty-2的CRD是被S-酰基转移酶差异修饰的。高特异性/低活性的zDHHC17酶介导的Sprouty-2的限制性S酰化，以及半胱氨酸-265和-268被确定为该酶的关键靶标。相反，低特异性/高活性的zDHHC3和zDHHC7酶介导了更广泛的Sprouty-2 CRD修饰。然而，所有酶的S酰化都促进了Sprouty-2的表达，表明S酰化稳定了该蛋白。此外，我们还鉴定了两个电荷残基(天冬氨酸-214和赖氨酸-223)，它们存在于CRD中预测的α-螺旋的相反面上，这对于Sprouty-2的S酰化是必不可少的。有趣的是，干扰S酰化的突变也导致了Sprouty-2在PC12细胞中质膜定位的丢失。本研究揭示了Sprouty-2 S酰化反应的机制和结果，并强调了不同类型的zDHHC酶介导的S酰化反应的不同模式。肿瘤抑制蛋白Sprouty-2的S酰化需要特定的半胱氨酸和富含半胱氨酸的区域的带电残基，并调节质膜靶向。
Sprouty-2 is an important regulator of growth factor signalling and a tumour suppressor protein. The defining feature of this protein is a cysteine-rich domain (CRD) that contains twenty-six cysteine residues and is modified by S-acylation. In this study, we show that the CRD of sprouty-2 is differentially modified by S-acyltransferase enzymes. The high specificity/low activity zDHHC17 enzyme mediated restricted S-acylation of sprouty-2, and cysteine-265 and -268 were identified as key targets of this enzyme. In contrast, the low specificity/high activity zDHHC3 and zDHHC7 enzymes mediated more expansive modification of the sprouty-2 CRD. Nevertheless, S-acylation by all enzymes enhanced sprouty-2 expression, suggesting that S-acylation stabilises this protein. In addition, we identified two charged residues (aspartate-214 and lysine-223), present on opposite faces of a predicted α-helix in the CRD, which are essential for S-acylation of sprouty-2. Interestingly, mutations that perturbed S-acylation also led to a loss of plasma membrane localisation of sprouty-2 in PC12 cells. This study provides insight into the mechanisms and outcomes of sprouty-2 S-acylation, and highlights distinct patterns of S-acylation mediated by different classes of zDHHC enzymes. S-acylation of the tumour suppressor protein sprouty-2 requires specific cysteine and charged residues in the cysteine-rich domain, and regulates plasma membrane targeting.