Xuepeng Wei, Kollin Schultz, Gleb A. Bazilevsky, Austin D. Vogt, R. Marmorstein
{"title":"Structure of ACLY in complex with CoA","authors":"Xuepeng Wei, Kollin Schultz, Gleb A. Bazilevsky, Austin D. Vogt, R. Marmorstein","doi":"10.2210/pdb6poe/pdb","DOIUrl":null,"url":null,"abstract":"ATP-citrate lyase (ACLY) synthesizes cytosolic acetyl coenzyme A (acetyl-CoA), a fundamental cellular building block. Accordingly, aberrant ACLY activity is observed in many diseases. Here we report cryo-EM structures of human ACLY, alone or bound to substrates or products. ACLY forms a homotetramer with a rigid citrate synthase homology (CSH) module, flanked by four flexible acetyl-CoA synthetase homology (ASH) domains; CoA is bound at the CSH-ASH interface in mutually exclusive productive or unproductive conformations. The structure of a catalytic mutant of ACLY in the presence of ATP, citrate and CoA substrates reveals a phospho-citryl-CoA intermediate in the ASH domain. ACLY with acetyl-CoA and oxaloacetate products shows the products bound in the ASH domain, with an additional oxaloacetate in the CSH domain, which could function in ACLY autoinhibition. These structures, which are supported by biochemical and biophysical data, challenge previous proposals of the ACLY catalytic mechanism and suggest additional therapeutic possibilities for ACLY-associated metabolic disorders.","PeriodicalId":18836,"journal":{"name":"Nature Structural &Molecular Biology","volume":null,"pages":null},"PeriodicalIF":16.8000,"publicationDate":"2019-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nature Structural &Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2210/pdb6poe/pdb","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
ATP-citrate lyase (ACLY) synthesizes cytosolic acetyl coenzyme A (acetyl-CoA), a fundamental cellular building block. Accordingly, aberrant ACLY activity is observed in many diseases. Here we report cryo-EM structures of human ACLY, alone or bound to substrates or products. ACLY forms a homotetramer with a rigid citrate synthase homology (CSH) module, flanked by four flexible acetyl-CoA synthetase homology (ASH) domains; CoA is bound at the CSH-ASH interface in mutually exclusive productive or unproductive conformations. The structure of a catalytic mutant of ACLY in the presence of ATP, citrate and CoA substrates reveals a phospho-citryl-CoA intermediate in the ASH domain. ACLY with acetyl-CoA and oxaloacetate products shows the products bound in the ASH domain, with an additional oxaloacetate in the CSH domain, which could function in ACLY autoinhibition. These structures, which are supported by biochemical and biophysical data, challenge previous proposals of the ACLY catalytic mechanism and suggest additional therapeutic possibilities for ACLY-associated metabolic disorders.
atp -柠檬酸解酶(ACLY)合成胞浆乙酰辅酶A (acetyl- coa),这是细胞的基本组成部分。因此,在许多疾病中观察到异常的ACLY活性。在这里,我们报告了人类ACLY的低温电镜结构,单独或结合底物或产物。ACLY形成一个具有刚性柠檬酸合成酶同源性(CSH)模块的同聚体,两侧是四个柔性乙酰辅酶a合成酶同源性(ASH)结构域;CoA以互斥的生产性或非生产性构象结合在CSH-ASH界面上。在ATP、柠檬酸盐和CoA底物存在下,ACLY的催化突变体的结构揭示了ASH域中的磷酸柠檬酸辅酶a中间体。含有乙酰辅酶a和草酰乙酸产物的ACLY表明产物结合在ASH结构域,在CSH结构域有一个额外的草酰乙酸,这可能在ACLY中起自抑制作用。这些结构得到了生化和生物物理数据的支持,挑战了之前关于ACLY催化机制的建议,并为ACLY相关代谢疾病的治疗提供了额外的可能性。
期刊介绍:
Nature Structural & Molecular Biology is a monthly journal that focuses on the functional and mechanistic understanding of how molecular components in a biological process work together. It serves as an integrated forum for structural and molecular studies. The journal places a strong emphasis on the functional and mechanistic understanding of how molecular components in a biological process work together. Some specific areas of interest include the structure and function of proteins, nucleic acids, and other macromolecules, DNA replication, repair and recombination, transcription, regulation of transcription and translation, protein folding, processing and degradation, signal transduction, and intracellular signaling.