S-acylation of Sprouty and SPRED proteins by the S-acyltransferase zDHHC17 involves a novel mode of enzyme-substrate interaction.
S-acylation of Sprouty and SPRED proteins by the S-acyltransferase zDHHC17 involves a novel mode of enzyme-substrate interaction.
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DOI：
10.1016/j.jbc.2022.102754
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发表时间：
2023-01
期刊：
影响因子：
--
通讯作者：
Chamberlain LH
中科院分区：
文献类型：
--
作者：
Butler L;Locatelli C;Allagioti D;Lousa I;Lemonidis K;Tomkinson NCO;Salaun C;Chamberlain LH
S-酰化是一种重要的翻译后修饰，其由人类中的23种zDHHC酶家族介导。数千种蛋白质被S-酰化修饰;然而，我们缺乏对如何实现酶-底物识别和特异性的详细了解。以前的工作表明，锚蛋白重复结构域的zDHHC 17（ANK 17）识别短的线性基序，称为zDHHC ANK结合基序（zDABM）在底物蛋白SNAP 25，作为一种机制，底物招募之前的S-酰化。在这里，我们研究了zDHHC 17对Sprouty和SPRED家族蛋白质的S-酰化。有趣的是，尽管Sprouty-2（Spry 2）含有与ANK 17相互作用的zDABM，但这种结合模式对于S-酰化是不合适的，并且实际上去除zDABM并不完全消除与zDHHC 17的结合。此外，相关的SPRED 3蛋白与zDHHC 17相互作用并被zDHHC 17有效地S-酰化，尽管缺乏zDABM。我们对SPRED 3进行了突变分析，以更好地了解其与zDHHC 17的zDABM非依赖性相互作用的基础。该分析发现，SPRED 3的富含半胱氨酸的SPR结构域（其是所有Sprouty和SPRED蛋白的定义特征）与zDHHC 17相互作用。令人惊讶的是，与SPRED 3的相互作用不依赖于ANK 17。我们对Spry 2的突变分析与含有zDHHC 17结合位点的该蛋白的SPR结构域一致，并且Spry 2还显示出与缺乏ANK结构域的zDHHC 17突变体的可检测结合。因此，zDHHC 17可以通过zDABM依赖性和/或zDABM非依赖性机制识别其底物，并且一些底物显示出与该酶结合的一种以上模式。
S-acylation is an essential post-translational modification, which is mediated by a family of 23 zDHHC enzymes in humans. Several thousand proteins are modified by S-acylation; however, we lack a detailed understanding of how enzyme–substrate recognition and specificity is achieved. Previous work showed that the ankyrin repeat domain of zDHHC17 (ANK17) recognizes a short linear motif, known as the zDHHC ANK binding motif (zDABM) in substrate protein SNAP25, as a mechanism of substrate recruitment prior to S-acylation. Here, we investigated the S-acylation of the Sprouty and SPRED family of proteins by zDHHC17. Interestingly, although Sprouty-2 (Spry2) contains a zDABM that interacts with ANK17, this mode of binding is dispensable for S-acylation, and indeed removal of the zDABM does not completely ablate binding to zDHHC17. Furthermore, the related SPRED3 protein interacts with and is efficiently S-acylated by zDHHC17, despite lacking a zDABM. We undertook mutational analysis of SPRED3 to better understand the basis of its zDABM-independent interaction with zDHHC17. This analysis found that the cysteine-rich SPR domain of SPRED3, which is the defining feature of all Sprouty and SPRED proteins, interacts with zDHHC17. Surprisingly, the interaction with SPRED3 was independent of ANK17. Our mutational analysis of Spry2 was consistent with the SPR domain of this protein containing a zDHHC17-binding site, and Spry2 also showed detectable binding to a zDHHC17 mutant lacking the ANK domain. Thus, zDHHC17 can recognize its substrates through zDABM-dependent and/or zDABM–independent mechanisms, and some substrates display more than one mode of binding to this enzyme.
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DOI：
10.1126/science.aao6326
发表时间：
2018-01-12
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Rana MS;Kumar P;Lee CJ;Verardi R;Rajashankar KR;Banerjee A
通讯作者：
Banerjee A
DOI：
10.1074/jbc.m117.799650
发表时间：
2017-10-20
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lemonidis K;MacLeod R;Baillie GS;Chamberlain LH
通讯作者：
Chamberlain LH
DOI：
10.1074/jbc.m802140200
发表时间：
2008-09-05
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Greaves J;Salaun C;Fukata Y;Fukata M;Chamberlain LH
通讯作者：
Chamberlain LH
影响因子：
4.8
作者：
Lobo, S;Greentree, WK;Deschenes, RJ
通讯作者：
Deschenes, RJ
影响因子：
4
作者：
Locatelli C;Lemonidis K;Salaun C;Tomkinson NCO;Chamberlain LH
通讯作者：
Chamberlain LH