The S-acyltransferase zDHHC2 mediates dynamic S-acylation of PSD95 and AKAP79/150, which impacts synaptic targeting of AMPA receptors. zDHHC2 is responsive to synaptic activity and catalyses the increased S-acylation of PSD95 that occurs following action potential blockade or application of ionotropic glutamate receptor antagonists. These treatments have been proposed to increase plasma membrane delivery of zDHHC2 via an endosomal cycling pathway, enhancing substrate accessibility. To generate an improved understanding of zDHHC2 trafficking and how this might be regulated by neuronal activity, we searched for intramolecular signals that regulate enzyme localisation. Two signals were mapped to the C-terminal tail of zDHHC2: a non-canonical dileucine motif [SxxxLL] and a downstream NP motif. Mutation of these signals enhanced plasma membrane accumulation of zDHHC2 in both neuroendocrine PC12 cells and rat hippocampal neurons, consistent with reduced endocytic retrieval. Furthermore, mutation of these signals also increased accumulation of the enzyme in neurites. Interestingly, several threonine and serine residues are adjacent to these sorting motifs and analysis of phospho-mimetic mutants highlighted a potential role for phosphorylation in regulating the efficacy of these signals. This study offers new molecular insight into the signals that determine zDHHC2 localisation and highlights a potential mechanism to regulate these trafficking signals.

S-酰基转移酶zDHHC2介导PSD95和AKAP79 / 150的动态S-酰化，这影响AMPA受体的突触靶向。zDHHC2对突触活性有反应，并催化PSD95的增加的S-酰化作用，该作用在动作电位阻断或应用离子型谷氨酸受体拮抗剂后发生。已经提出这些治疗以增加经由内体循环途径的zDHHC2的质膜递送，增强底物可及性。为了更好地了解zDHHC2的转运以及如何通过神经元活性调节它，我们搜索了调节酶定位的分子内信号。两个信号被映射到zDHHC2的C末端尾部：一个非典型的双亮氨酸基序[SxxxLL]和一个下游NP基序。这些信号的突变增强了神经内分泌PC12细胞和大鼠海马神经元中zDHHC2的质膜积累，这与减少的内吞恢复一致。此外，这些信号的突变也增加了神经突中酶的积累。有趣的是，一些苏氨酸和丝氨酸残基与这些排序基序相邻，并且模拟磷酸突变体的分析突出了磷酸化在调节这些信号的效力中的潜在作用。这项研究为确定zDHHC2定位的信号提供了新的分子见解，并强调了调节这些运输信号的潜在机制。这些信号的突变也增加了神经突中酶的积累。有趣的是，一些苏氨酸和丝氨酸残基与这些排序基序相邻，并且模拟磷酸突变体的分析突出了磷酸化在调节这些信号的效力中的潜在作用。这项研究为确定zDHHC2定位的信号提供了新的分子见解，并强调了调节这些运输信号的潜在机制。这些信号的突变也增加了神经突中酶的积累。有趣的是，一些苏氨酸和丝氨酸残基与这些排序基序相邻，并且模拟磷酸突变体的分析突出了磷酸化在调节这些信号的效力中的潜在作用。这项研究为确定zDHHC2定位的信号提供了新的分子见解，并强调了调节这些运输信号的潜在机制。