重组小鼠成釉蛋白在大肠杆菌中的表达与纯化
The Expression and Purification of Recombinant Mouse Ameloblastin in E. coli.
作者信息
Su Jingtan, Bapat Rucha Arun, Moradian-Oldak Janet
机构信息
Center for Craniofacial Molecular Biology, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA, USA.
Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, USA.
出版信息
Methods Mol Biol. 2019;1922:229-236. doi: 10.1007/978-1-4939-9012-2_23.
Abstract
Ameloblastin is the second most abundant enamel matrix protein, and is thought to be essential for ameloblast cell polarization, cell adhesion, and enamel mineralization. However, studies of ameloblastin's function and its molecular mechanism have been limited due to difficulty in obtaining recombinant ameloblastin in vitro. Here, we present a protocol for successful ameloblastin expression and purification in E. coli.
摘要
成釉蛋白是牙釉质基质中含量第二丰富的蛋白质，被认为对成釉细胞的极化、细胞黏附以及牙釉质矿化至关重要。然而，由于在体外难以获得重组成釉蛋白，关于成釉蛋白功能及其分子机制的研究一直有限。在此，我们介绍一种在大肠杆菌中成功表达和纯化成釉蛋白的方法。
相似文献
1
The Expression and Purification of Recombinant Mouse Ameloblastin in E. coli.重组小鼠成釉蛋白在大肠杆菌中的表达与纯化
Methods Mol Biol. 2019;1922:229-236. doi: 10.1007/978-1-4939-9012-2_23.
2
Full-length sequence, localization, and chromosomal mapping of ameloblastin. A novel tooth-specific gene.成釉蛋白的全长序列、定位及染色体图谱。一个新的牙齿特异性基因。
J Biol Chem. 1996 Feb 23;271(8):4431-5. doi: 10.1074/jbc.271.8.4431.
3
Protein Interaction between Ameloblastin and Proteasome Subunit α Type 3 Can Facilitate Redistribution of Ameloblastin Domains within Forming Enamel.成釉蛋白与蛋白酶体亚基α3型之间的蛋白质相互作用可促进成釉蛋白结构域在形成的釉质内重新分布。
J Biol Chem. 2015 Aug 21;290(34):20661-20673. doi: 10.1074/jbc.M115.640185. Epub 2015 Jun 12.
4
Essential roles of ameloblastin in maintaining ameloblast differentiation and enamel formation.成釉蛋白在维持成釉细胞分化和釉质形成中的重要作用。
Cells Tissues Organs. 2005;181(3-4):189-95. doi: 10.1159/000091380.
5
Escherichia coli peptide methionine sulfoxide reductase: cloning, high expression, and purification.大肠杆菌肽甲硫氨酸亚砜还原酶：克隆、高表达及纯化
Methods Enzymol. 1995;251:462-70. doi: 10.1016/0076-6879(95)51150-4.
6
Interaction between the enamel matrix proteins amelogenin and ameloblastin.釉基质蛋白釉原蛋白和成釉蛋白之间的相互作用。
Biochem Biophys Res Commun. 2004 Oct 22;323(3):1075-83. doi: 10.1016/j.bbrc.2004.08.207.
7
Intrinsically disordered enamel matrix protein ameloblastin forms ribbon-like supramolecular structures via an N-terminal segment encoded by exon 5.无规则卷曲的釉基质蛋白牙釉蛋白通过由外显子 5 编码的 N 端片段形成带状超分子结构。
J Biol Chem. 2013 Aug 2;288(31):22333-45. doi: 10.1074/jbc.M113.456012. Epub 2013 Jun 19.
8
High-Throughput Production of Proteins in E. coli for Structural Studies.用于结构研究的大肠杆菌中蛋白质的高通量生产。
Methods Mol Biol. 2017;1586:359-371. doi: 10.1007/978-1-4939-6887-9_24.
9
Purification of recombinant Gi alpha and Go alpha proteins from Escherichia coli.从大肠杆菌中纯化重组Giα和Goα蛋白。
Methods Enzymol. 1991;195:202-15. doi: 10.1016/0076-6879(91)95167-i.
10
Recombinant human enamelin produced in Escherichia coli promotes mineralization in vitro.重组大肠杆菌来源的人釉蛋白促进体外矿化。
BMC Biotechnol. 2024 Jul 9;24(1):48. doi: 10.1186/s12896-024-00875-0.
引用本文的文献
1
The Dynamic Interactions of a Multitargeting Domain in Ameloblastin Protein with Amelogenin and Membrane.牙釉蛋白中多靶向结构域与釉原蛋白和细胞膜的动态相互作用
Int J Mol Sci. 2023 Feb 9;24(4):3484. doi: 10.3390/ijms24043484.
2
Early evolution of enamel matrix proteins is reflected by pleiotropy of physiological functions.牙釉质基质蛋白的早期进化反映了其生理功能的多效性。
Sci Rep. 2023 Jan 26;13(1):1471. doi: 10.1038/s41598-023-28388-4.
3
Modeling ameloblast-matrix interactions using 3D cell culture.使用三维细胞培养模拟成釉细胞与基质的相互作用。
Front Physiol. 2022 Dec 1;13:1069519. doi: 10.3389/fphys.2022.1069519. eCollection 2022.
4
Coemergence of the Amphipathic Helix on Ameloblastin With Mammalian Prismatic Enamel.牙釉蛋白与哺乳动物棱柱形釉质中两亲螺旋的共出现。
Mol Biol Evol. 2022 Nov 3;39(11). doi: 10.1093/molbev/msac205.
5
Data from ameloblast cell lines cultured in 3D using various gel substrates in the presence of ameloblastin.在成釉蛋白存在的情况下，使用各种凝胶底物在三维环境中培养的成釉细胞系的数据。
Data Brief. 2022 May 5;42:108233. doi: 10.1016/j.dib.2022.108233. eCollection 2022 Jun.
6
Regulation of Hydroxyapatite Nucleation In Vitro through Ameloblastin-Amelogenin Interactions.通过釉原蛋白-釉基质蛋白相互作用调控体外羟基磷灰石成核。
ACS Biomater Sci Eng. 2023 Apr 10;9(4):1834-1842. doi: 10.1021/acsbiomaterials.1c01113. Epub 2022 Jan 24.
7
Ameloblastin promotes polarization of ameloblast cell lines in a 3-D cell culture system.成釉蛋白促进三维细胞培养体系中成釉细胞瘤细胞系的极化。
Matrix Biol. 2022 Jan;105:72-86. doi: 10.1016/j.matbio.2021.11.002. Epub 2021 Nov 20.
8
Co-Immunoprecipitation Reveals Interactions Between Amelogenin and Ameloblastin Their Self-Assembly Domains.免疫共沉淀揭示了釉原蛋白和成釉蛋白及其自组装结构域之间的相互作用。
Front Physiol. 2020 Dec 23;11:622086. doi: 10.3389/fphys.2020.622086. eCollection 2020.
9
An Evolutionarily Conserved Helix Mediates Ameloblastin-Cell Interaction.一个进化上保守的螺旋介导釉原蛋白与细胞的相互作用。
J Dent Res. 2020 Aug;99(9):1072-1081. doi: 10.1177/0022034520918521. Epub 2020 May 13.
10
Ameloblastin Binds to Phospholipid Bilayers via a Helix-Forming Motif within the Sequence Encoded by Exon 5.成釉蛋白通过外显子5编码序列内的螺旋形成基序与磷脂双层结合。
ACS Omega. 2019 Feb 28;4(2):4405-4416. doi: 10.1021/acsomega.8b03582.
本文引用的文献
1
Intrinsically disordered proteins drive enamel formation via an evolutionarily conserved self-assembly motif.内在无序蛋白质通过一种进化保守的自组装基序驱动牙釉质形成。
Proc Natl Acad Sci U S A. 2017 Feb 28;114(9):E1641-E1650. doi: 10.1073/pnas.1615334114. Epub 2017 Feb 14.
2
The Importance of Serine Phosphorylation of Ameloblastin on Enamel Formation.成釉蛋白丝氨酸磷酸化在釉质形成中的重要性。
J Dent Res. 2016 Nov;95(12):1408-1414. doi: 10.1177/0022034516661513. Epub 2016 Jul 29.
3
Evolutionary analysis of selective constraints identifies ameloblastin (AMBN) as a potential candidate for amelogenesis imperfecta.选择性限制的进化分析确定釉成熟蛋白（AMBN）为牙釉质发育不全的一个潜在候选因素。
BMC Evol Biol. 2015 Jul 30;15:148. doi: 10.1186/s12862-015-0431-0.
4
Deletion of ameloblastin exon 6 is associated with amelogenesis imperfecta.成釉蛋白外显子6的缺失与牙釉质发育不全相关。
Hum Mol Genet. 2014 Oct 15;23(20):5317-24. doi: 10.1093/hmg/ddu247. Epub 2014 May 23.
5
Protein-mediated enamel mineralization.蛋白质介导的牙釉质矿化。
Front Biosci (Landmark Ed). 2012 Jun 1;17(6):1996-2023. doi: 10.2741/4034.
6
Cleavage site specificity of MMP-20 for secretory-stage ameloblastin.MMP-20 对分泌期釉原蛋白的酶切位点特异性。
J Dent Res. 2010 Aug;89(8):785-90. doi: 10.1177/0022034510366903. Epub 2010 Apr 16.
7
Bioinformatic analysis and molecular modelling of human ameloblastin suggest a two-domain intrinsically unstructured calcium-binding protein.人釉原蛋白的生物信息学分析和分子建模表明其为一种双结构域内在无序的钙结合蛋白。
Eur J Oral Sci. 2008 Apr;116(2):124-34. doi: 10.1111/j.1600-0722.2008.00526.x.
8
Amelogenin and ameloblastin show growth-factor like activity in periodontal ligament cells.釉原蛋白和成釉蛋白在牙周膜细胞中表现出类生长因子活性。
Eur J Oral Sci. 2006 May;114 Suppl 1:244-53; discussion 254-6, 381-2. doi: 10.1111/j.1600-0722.2006.00322.x.
9
Ameloblastin is a cell adhesion molecule required for maintaining the differentiation state of ameloblasts.成釉蛋白是维持成釉细胞分化状态所需的一种细胞黏附分子。
J Cell Biol. 2004 Dec 6;167(5):973-83. doi: 10.1083/jcb.200409077.
10
Interaction between the enamel matrix proteins amelogenin and ameloblastin.釉基质蛋白釉原蛋白和成釉蛋白之间的相互作用。
Biochem Biophys Res Commun. 2004 Oct 22;323(3):1075-83. doi: 10.1016/j.bbrc.2004.08.207.
Zotero 插件