Peptide array-based screening reveals a large number of proteins interacting with the ankyrin-repeat domain of the zDHHC17 S-acyltransferase.
Peptide array-based screening reveals a large number of proteins interacting with the ankyrin-repeat domain of the zDHHC17 S-acyltransferase.
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DOI：
10.1074/jbc.m117.799650
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发表时间：
2017-10-20
期刊：
影响因子：
--
通讯作者：
Chamberlain LH
中科院分区：
文献类型：
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作者：
Lemonidis K;MacLeod R;Baillie GS;Chamberlain LH
zDHHC S-酰基转移酶是催化蛋白S-酰化的酶，S-酰化是蛋白上常见的翻译后修饰，经常影响其膜靶向和运输。zDHHC 17（HIP 14）和zDHHC 13（HIP 14 L）S-酰基转移酶的锚蛋白重复（AR）结构域参与底物募集和S-酰化独立功能，最近显示通过特异性识别其中的共有序列结合至少六种蛋白质。为了进一步完善与zDHHC 17 AR结合的规则，我们采用了基于突触体相关蛋白25（SNAP 25）和半胱氨酸串蛋白α（CSPα）的zDHHC AR结合基序（zDABM）序列的肽阵列。400肽的结合偏好的定量比较使我们能够构建zDHHC 17 AR结合的位置特异性评分矩阵（PSSM），我们预测并随后验证了许多推定的zDHHC 17相互作用。我们鉴定了95个人zDABM序列，这些序列在氨基酸使用中具有意想不到的多功能性;这些序列分布在90种蛋白质中，其中62种先前未涉及zDHHC 17/13结合。这些含zDABM的蛋白质包括SNAP 25、sprougty、cornifelin、锚蛋白和含SLAIN基序的家族的所有家族成员;共享相同结合序列的七种内源性Gag多聚蛋白;以及参与细胞骨架组织、细胞通讯和信号传导调节的几种蛋白质。一打含zDABM的蛋白质具有多于一个的zDABM序列，而对于Ena/VASP样蛋白质，鉴定了与zDHHC 17的AR的同种型特异性结合。人类蛋白质组中大量的zDABM序列表明zDHHC 17可能是调节许多细胞过程的相互作用中心。
zDHHC S-acyltransferases are enzymes catalyzing protein S-acylation, a common post-translational modification on proteins frequently affecting their membrane targeting and trafficking. The ankyrin repeat (AR) domain of zDHHC17 (HIP14) and zDHHC13 (HIP14L) S-acyltransferases, which is involved in both substrate recruitment and S-acylation-independent functions, was recently shown to bind at least six proteins, by specific recognition of a consensus sequence in them. To further refine the rules governing binding to the AR of zDHHC17, we employed peptide arrays based on zDHHC AR-binding motif (zDABM) sequences of synaptosomal-associated protein 25 (SNAP25) and cysteine string protein α (CSPα). Quantitative comparisons of the binding preferences of 400 peptides allowed us to construct a position-specific scoring matrix (PSSM) for zDHHC17 AR binding, with which we predicted and subsequently validated many putative zDHHC17 interactors. We identified 95 human zDABM sequences with unexpected versatility in amino acid usage; these sequences were distributed among 90 proteins, of which 62 have not been previously implicated in zDHHC17/13 binding. These zDABM-containing proteins included all family members of the SNAP25, sprouty, cornifelin, ankyrin, and SLAIN-motif containing families; seven endogenous Gag polyproteins sharing the same binding sequence; and several proteins involved in cytoskeletal organization, cell communication, and regulation of signaling. A dozen of the zDABM-containing proteins had more than one zDABM sequence, whereas isoform-specific binding to the AR of zDHHC17 was identified for the Ena/VASP-like protein. The large number of zDABM sequences within the human proteome suggests that zDHHC17 may be an interaction hub regulating many cellular processes.