Regulation of Calcium Signaling by Protein Lipidation

蛋白质脂化对钙信号传导的调节

• 批准号: 10404120
• 负责人: Askar Akimzhanov
• 金额: $ 33.27万
• 依托单位: ROWAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10404120
• 关键词: Acylation; Acyltransferase; Affect; Agonist; Animals; Biochemical; Biological; Calcium; Calcium Channel; Calcium Signaling; Calcium ion; Calcium-Binding Domain; Caveolae; Cell membrane; Cell physiology; Cells; Clinical Data; Complex; Cysteine; Cytoplasm; Data; Dependence; Disease; Enzymes; Family; Genetic study; Homeostasis; Immune response; Immune system; Immunologic Deficiency Syndromes; Ion Channel; Jurkat Cells; Kinetics; Label; Life Cycle Stages; Lipids; Mediating; Membrane; Membrane Microdomains; Memory; Metabolic; Methods; Monitor; Mutation; Plant Resins; Play; Post-Translational Protein Processing; Process; Proteins; Proteomics; Receptor Activation; Regulation; Role; STIM1 gene; Second Messenger Systems; Signal Transduction; Signaling Protein; Stimulus; Syndrome; T cell regulation; T-Cell Activation; T-Cell Lymphoma; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Total Internal Reflection Fluorescent; Tubular Aggregate Myopathies; congenital immunodeficiency; enzyme activity; experimental study; human disease; immunological synapse; in vivo; long chain fatty acid; loss of function mutation; mutant; novel; protein acyltransferase; recruit; response; therapeutic target; trafficking

ABSTRACT Intracellular calcium signals play a vital role in regulating immune system homeostasis and function. In T cells, calcium ions serve as a critical second messenger in a broad variety of cellular processes regulating T cell activation, proliferation and differentiation of naïve T cells into effector or memory cells. The mechanism supporting the sustained calcium influx into the cytoplasm is known as store-operated calcium entry (SOCE). Two proteins, Orai1 and STIM1, were identified as primary modulators of SOCE in T cells. SOCE is initiated when STIM1 senses the depletion of internal calcium stores and associates with the pore-forming Orai1 to assemble the calcium release-activated channel. The critical role of STIM1 and Orai1 in the regulation of T cell immune responses is well supported by genetic studies performed in animals as well as clinical data. Biological consequences of Orai1 or STIM1 deficiencies include severe immunodeficiency, tubular aggregate myopathy, and Stormorken syndrome. In our preliminary experiments, we have identified both Orai1 and STIM1 as endogenously S-acylated proteins. S-acylation, a reversible post-translational lipidation of cysteine residues with long-chain fatty acids, is catalyzed by the family of DHHC palmitoyl acyltransferases known to regulate the function of many key T cell signaling proteins. Our previous studies strongly suggest that stimulus-dependent protein lipidation is an essential part of the intricate signaling machinery controlling T cell activation and function. Therefore, we hypothesize that dynamic S-acylation of Orai1 and STIM1 is a critical regulator of calcium entry in T cells. To uncover the role of protein lipidation in calcium signaling, we will (1) determine whether Orai1 and STIM1 are S-acylated proteins in T cells, (2) determine the functional consequences of Orai1 and STIM1 acylation and (3) identify the enzymatic mechanisms mediating Orai1 and STIM1 S-acylation. The successful completion of the proposed project will demonstrate the biological significance of protein lipidation in regulation of SOCE as well as the role of palmitoyl acyltransferases in regulation of the calcium signaling in T cells with relevance to primary immunodeficiency disease.

摘要 细胞内钙信号在调节免疫系统的动态平衡和功能方面起着至关重要的作用。在T细胞中， 钙离子在调节T细胞的多种细胞过程中起着关键的第二信使作用 幼稚T细胞的激活、增殖和分化为效应细胞或记忆细胞。这一机制 支持钙离子持续内流到细胞质称为储备型钙内流(SOCE)。 Orai1和STIM1是T细胞SOCE的主要调节因子。启动SOCE 当STIM1感觉到内部钙库的耗尽并与成孔Orai1联系在一起时 组装钙释放激活通道。STIM1和Orai1在T细胞调节中的关键作用 在动物身上进行的遗传研究和临床数据很好地支持了免疫反应。生物学 Orai1或STIM1缺陷的后果包括严重的免疫缺陷，管状聚集性肌病， 和斯托莫肯综合征。在我们的初步实验中，我们已经确定Orai1和STIM1都是 内源性S酰化蛋白。S-酰化，半胱氨酸残基的可逆翻译后脂肪基化 与长链脂肪酸，是由DHHC棕榈酰基转移酶家族催化的，该家族调节 许多关键的T细胞信号蛋白的功能。我们之前的研究强烈表明，依赖刺激 蛋白质脂化是控制T细胞激活和释放的复杂信号机制的重要组成部分。 功能。因此，我们推测Orai1和STIM1的动态S酰化是一个关键的调节因子 T细胞内钙离子的进入。为了揭示蛋白质脂化在钙信号中的作用，我们将(1)确定 Orai1和STim1是否是T细胞中的S酰化蛋白，(2)决定了 ORAI1和STIM1的酰化反应；(3)确定了OraI1和STIM1的S酰化反应的酶机制。 拟议项目的成功完成将证明蛋白质的生物学意义。 脂肪在SOCE调节中的作用及棕榈酰基转移酶在钙调节中的作用 T细胞中的信号与原发免疫缺陷疾病相关。

项目成果

Regulation of Dynamic Protein S-Acylation.

• DOI: 10.3389/fmolb.2021.656440
• 发表时间: 2021
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Chen JJ;Fan Y;Boehning D
• 通讯作者: Boehning D

S-acylation of STIM1 regulates store-operated calcium entry.

STIM1 的 S-酰化调节钙库操作的钙进入。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Kodakandla,Goutham;West,Savannah;Wang,Qioachu;Tewari,Ritika;Zhu,Michael;Akimzhanov,Askar;Boehning,DarrenF
• 通讯作者: Boehning,DarrenF