Phosphoproteomic and proteomic profiling of serine/threonine protein kinase PkaE of <i>Streptomyces coelicolor</i> A3(2) and its role in secondary metabolism and morphogenesis

This study aimed to investigate the role of serine/threonine kinase PkaE in <i>Streptomyces coelicolor</i> A3(2). Liquid chromatography tandem mass spectrometry was performed for comparative phosphoproteome and proteome analyses of <i>S. coelicolor</i> A3(2), followed by an <i>in vitro</i> phosphorylation assay. Actinorhodin production in the <i>pkaE</i> deletion mutant was lower than that in wild-type <i>S. coelicolor</i> A3(2), and the spores of the <i>pkaE</i> deletion mutant were damaged. Furthermore, phosphoproteome analysis revealed that 6 proteins were significantly differentially hypophosphorylated in <i>pkaE</i> deletion mutant (<i>p</i> in vitro phosphorylation assay revealed that PkaE phosphorylated FtsZ. Comparative proteome analysis revealed 362 differentially expressed proteins (<i>p</i> pkaE deletion mutant involved in actinorhodin biosynthesis. Gene ontology enrichment analysis revealed that PkaE participates in various biological and cellular processes. Hence, <i>S. coelicolor</i> PkaE participates in actinorhodin biosynthesis and morphogenesis. Liquid chromatography tandem mass spectrometry was performed for comparative phosphoproteome and proteome analyses of <i>S. coelicolor</i> A3(2) wild-type and <i>pkaE</i> deletion mutant, followed by an <i>in vitro</i> phosphorylation assay.

本研究旨在探究丝氨酸/苏氨酸激酶PkaE在天蓝色链霉菌（Streptomyces coelicolor）A3(2)中的作用。研究采用液相色谱-串联质谱（liquid chromatography tandem mass spectrometry），对天蓝色链霉菌A3(2)的磷酸化蛋白质组与蛋白质组开展比较分析，并辅以体外磷酸化实验。pkaE缺失突变体的放线紫红素产量低于野生型天蓝色链霉菌A3(2)，且该缺失突变体的孢子形态受损。磷酸化蛋白质组分析显示，pkaE缺失突变体中共有6种蛋白质呈现显著的差异低磷酸化；体外磷酸化实验证实，PkaE可磷酸化FtsZ。比较蛋白质组分析显示，pkaE缺失突变体中存在362种差异表达蛋白质，这些蛋白质参与放线紫红素的生物合成过程。基因本体（Gene Ontology）富集分析结果表明，PkaE参与多种生物学与细胞过程。综上，天蓝色链霉菌PkaE参与放线紫红素的生物合成及形态发生。本研究通过液相色谱-串联质谱，对野生型天蓝色链霉菌A3(2)与pkaE缺失突变体的磷酸化蛋白质组、蛋白质组进行比较分析，并辅以体外磷酸化实验。

搜集汇总

背景与挑战

背景概述

该数据集通过磷酸化蛋白质组学和蛋白质组学分析，研究了天蓝色链霉菌A3(2)中丝氨酸/苏氨酸蛋白激酶PkaE的功能。研究发现PkaE参与放线紫红素的生物合成和形态发生过程，其缺失会导致次级代谢产物减少和孢子损伤。分析揭示了多个差异磷酸化和表达蛋白，包括BldG和FtsZ，并通过体外实验证实PkaE能磷酸化FtsZ。