A proteomic approach identifies many novel palmitoylated proteins in Arabidopsis
A proteomic approach identifies many novel palmitoylated proteins in Arabidopsis
复制标题
DOI：
10.1111/nph.12077
复制
发表时间：
2013-02-01
期刊：
影响因子：
9.4
通讯作者：
Grierson, Claire S.
中科院分区：
文献类型：
--
作者：
Hemsley, Piers A.;Weimar, Thilo;Grierson, Claire S.
S-酰化（棕榈酰化）是蛋白质的翻译后修饰，涉及向半胱氨酸残基添加酰基脂质。S-酰化促进蛋白质与膜的结合，并影响蛋白质稳定性、微区分配、膜靶向和活化状态。不存在S-酰化的共有基序，因此需要经验鉴定。在这里，我们描述了一种基于生物素开关同量异位标记的相对和绝对定量（iTRAQ）方法来鉴定拟南芥中的S-酰化蛋白。我们使用这些数据来预测和确认不在我们数据集中的蛋白质的S-酰化。600种假定的S-酰化蛋白质影响不同的细胞过程。其中包括参与病原体感知和反应的蛋白质、有丝分裂原活化蛋白激酶（MAPKs）、富含亮氨酸重复受体样激酶（LRR-RLK）和RLK超家族成员、整体膜转运蛋白、ATP酶、可溶性N-乙基马来酰亚胺敏感因子活化蛋白受体（SNARE）和异源三聚体G蛋白。通过鉴定和确认SNARE和LRR-RLK家族中的S-酰化位点，证明了相关蛋白的S-酰化预测。我们发现，LRR-RLK FLS 2的S-酰化是鞭毛蛋白衍生肽flg 22诱导的完全反应所必需的，但不是定位到质膜所必需的。这些数据可用于鉴定相关蛋白质中的S-酰化位点。我们还证明了S-酰化是完整的LRR-RLK功能所必需的。
S-acylation (palmitoylation) is a poorly understood post-translational modification of proteins involving the addition of acyl lipids to cysteine residues. S-acylation promotes the association of proteins with membranes and influences protein stability, microdomain partitioning, membrane targeting and activation state. No consensus motif for S-acylation exists and it therefore requires empirical identification.Here, we describe a biotin switch isobaric tagging for relative and absolute quantification (iTRAQ)-based method to identify S-acylated proteins from Arabidopsis. We use these data to predict and confirm S-acylation of proteins not in our dataset.We identified c. 600 putative S-acylated proteins affecting diverse cellular processes. These included proteins involved in pathogen perception and response, mitogen-activated protein kinases (MAPKs), leucine-rich repeat receptor-like kinases (LRR-RLKs) and RLK superfamily members, integral membrane transporters, ATPases, soluble N-ethylmaleimide-sensitive factor-activating protein receptors (SNAREs) and heterotrimeric G-proteins. The prediction of S-acylation of related proteins was demonstrated by the identification and confirmation of S-acylation sites within the SNARE and LRR-RLK families. We showed that S-acylation of the LRR-RLK FLS2 is required for a full response to elicitation by the flagellin derived peptide flg22, but is not required for localization to the plasma membrane.Arabidopsis contains many more S-acylated proteins than previously thought. These data can be used to identify S-acylation sites in related proteins. We also demonstrated that S-acylation is required for full LRR-RLK function.