一种基于孔雀石绿的方法来评估葡聚糖磷酸酶活性。
A malachite green-based assay to assess glucan phosphatase activity.
机构信息
Department of Molecular and Cellular Biochemistry, Center for Structural Biology, University of Kentucky, Lexington, KY 40536, USA.
出版信息
Anal Biochem. 2013 Apr 1;435(1):54-6. doi: 10.1016/j.ab.2012.10.044. Epub 2012 Nov 29.
Abstract
With the recent discovery of a unique class of dual-specificity phosphatases that dephosphorylate glucans, we report an in vitro assay tailored for the detection of phosphatase activity against phosphorylated glucans. We demonstrate that, in contrast to a general phosphatase assay using a synthetic substrate, only phosphatases that possess glucan phosphatase activity liberate phosphate from the phosphorylated glucan amylopectin using the described assay. This assay is simple and cost-effective, providing reproducible results that clearly establish the presence or absence of glucan phosphatase activity. The assay described will be a useful tool in characterizing emerging members of the glucan phosphatase family.
摘要
最近发现了一类独特的双特异性磷酸酶，它们可以使葡聚糖去磷酸化，因此我们报告了一种针对磷酸化葡聚糖的磷酸酶活性检测的体外测定法。我们证明，与使用合成底物的一般磷酸酶测定法相反，只有具有葡聚糖磷酸酶活性的磷酸酶才能使用所述测定法从磷酸化的支链淀粉中释放出磷酸盐。该测定法简单且具有成本效益，可提供可重复的结果，明确确定是否存在葡聚糖磷酸酶活性。所描述的测定法将成为表征葡聚糖磷酸酶家族中新出现成员的有用工具。
相似文献
1
A malachite green-based assay to assess glucan phosphatase activity.一种基于孔雀石绿的方法来评估葡聚糖磷酸酶活性。
Anal Biochem. 2013 Apr 1;435(1):54-6. doi: 10.1016/j.ab.2012.10.044. Epub 2012 Nov 29.
2
Concanavalin A-Based Sedimentation Assay to Measure Substrate Binding of Glucan Phosphatases.基于伴刀豆球蛋白A的沉降试验来测量葡聚糖磷酸酶的底物结合情况。
J Vis Exp. 2022 Dec 23(190). doi: 10.3791/64700.
3
Cooperative Kinetics of the Glucan Phosphatase Starch Excess4.葡聚糖磷酸酶淀粉过量4的协同动力学
Biochemistry. 2021 Aug 10;60(31):2425-2435. doi: 10.1021/acs.biochem.1c00307. Epub 2021 Jul 28.
4
Structure of the Arabidopsis glucan phosphatase like sex four2 reveals a unique mechanism for starch dephosphorylation.拟南芥葡聚糖磷酸酶样蛋白 SEX42 的结构揭示了淀粉去磷酸化的独特机制。
Plant Cell. 2013 Jun;25(6):2302-14. doi: 10.1105/tpc.113.112706. Epub 2013 Jun 28.
5
Structural mechanisms of plant glucan phosphatases in starch metabolism.植物葡聚糖磷酸酶在淀粉代谢中的结构机制
FEBS J. 2016 Jul;283(13):2427-47. doi: 10.1111/febs.13703. Epub 2016 Mar 28.
6
The phosphoglucan phosphatase like sex Four2 dephosphorylates starch at the C3-position in Arabidopsis.磷酸葡聚糖磷酸酶样蛋白 SEX FOUR2 在拟南芥中于 C3 位去磷酸化淀粉。
Plant Cell. 2011 Nov;23(11):4096-111. doi: 10.1105/tpc.111.092155. Epub 2011 Nov 18.
7
Assessing the Biological Activity of the Glucan Phosphatase Laforin.评估葡聚糖磷酸酶拉佛林的生物活性。
Methods Mol Biol. 2016;1447:107-19. doi: 10.1007/978-1-4939-3746-2_7.
8
Mechanistic Insights into Glucan Phosphatase Activity against Polyglucan Substrates.针对多聚糖底物的葡聚糖磷酸酶活性的机制性见解
J Biol Chem. 2015 Sep 18;290(38):23361-70. doi: 10.1074/jbc.M115.658203. Epub 2015 Jul 31.
9
A bioassay for Lafora disease and laforin glucan phosphatase activity.拉福拉病和拉佛林葡聚糖磷酸酶活性的生物测定法。
Clin Biochem. 2013 Dec;46(18):1869-76. doi: 10.1016/j.clinbiochem.2013.08.016. Epub 2013 Sep 6.
10
Structural basis for the glucan phosphatase activity of Starch Excess4.淀粉过量 4 号的葡聚糖磷酸酶活性的结构基础。
Proc Natl Acad Sci U S A. 2010 Aug 31;107(35):15379-84. doi: 10.1073/pnas.1009386107. Epub 2010 Aug 2.
引用本文的文献
1
Structure and mechanism of biosynthesis of Streptococcus mutans cell wall polysaccharide.变形链球菌细胞壁多糖的生物合成结构与机制
Nat Commun. 2025 Jan 22;16(1):954. doi: 10.1038/s41467-025-56205-1.
2
2-Deoxy-4---inosose (DEI) is the Product of EboD, a Highly Conserved Dehydroquinate Synthase-like Enzyme in Bacteria and Eustigmatophyte Algae.2-脱氧-4---肌苷（DEI）是 EboD 的产物，EboD 是细菌和黄藻门真眼点藻中高度保守的脱氢奎宁酸合酶样酶。
ACS Chem Biol. 2024 Nov 15;19(11):2277-2283. doi: 10.1021/acschembio.4c00510. Epub 2024 Oct 15.
3
Catalytic Mechanism of Nonglycosidic C-N Bond Formation by the Pseudoglycosyltransferase Enzyme VldE.假糖基转移酶VldE催化非糖苷C-N键形成的机制
ACS Catal. 2023 Oct 20;13(20):13369-13382. doi: 10.1021/acscatal.3c02404. Epub 2023 Oct 3.
4
An empirical pipeline for personalized diagnosis of Lafora disease mutations.用于拉福拉病突变个性化诊断的经验性流程。
iScience. 2021 Oct 13;24(11):103276. doi: 10.1016/j.isci.2021.103276. eCollection 2021 Nov 19.
5
Generation and characterization of a laforin nanobody inhibitor.Laforin 纳米抗体抑制剂的产生和特性分析。
Clin Biochem. 2021 Jul;93:80-89. doi: 10.1016/j.clinbiochem.2021.03.017. Epub 2021 Apr 5.
6
Crystal structure of death-associated protein kinase 1 in complex with the dietary compound resveratrol.与膳食化合物白藜芦醇结合的死亡相关蛋白激酶1的晶体结构。
IUCrJ. 2021 Jan 1;8(Pt 1):131-138. doi: 10.1107/S2052252520015614.
7
The Pup-proteasome system regulates nitrate metabolism through an essential protein quality control pathway.Pup-proteasome 系统通过一种必需的蛋白质质量控制途径来调节硝酸盐代谢。
Proc Natl Acad Sci U S A. 2019 Feb 19;116(8):3202-3210. doi: 10.1073/pnas.1819468116. Epub 2019 Feb 5.
8
A novel EPM2A mutation yields a slow progression form of Lafora disease.一种新型的EPM2A突变导致拉福拉病的缓慢进展型。
Epilepsy Res. 2018 Sep;145:169-177. doi: 10.1016/j.eplepsyres.2018.07.003. Epub 2018 Jul 21.
9
Identification and analysis of OsttaDSP, a phosphoglucan phosphatase from Ostreococcus tauri.来自莱茵衣藻的磷酸葡聚糖磷酸酶OsttaDSP的鉴定与分析。
需注意，原文中的“Ostreococcus tauri”一般翻译为“莱茵衣藻” ，你提供的原文中该词有误，实际应该是“Ostreococcus tauri” ，我按照正确的物种名进行了翻译。 若按照你提供的错误物种名“Ostreococcus tauri” ，无法准确对应到已知的生物，翻译出来会比较奇怪，也不符合正常的学术内容。 你可根据实际情况进行调整。 以下是按照正确物种名翻译的最终译文：
来自莱茵衣藻的磷酸葡聚糖磷酸酶OsttaDSP的鉴定与分析。
PLoS One. 2018 Jan 23;13(1):e0191621. doi: 10.1371/journal.pone.0191621. eCollection 2018.
10
Interrogating Protein Phosphatases with Chemical Activity Probes.用具有化学活性的探针来探究蛋白磷酸酶。
Chemistry. 2018 Jun 4;24(31):7810-7824. doi: 10.1002/chem.201705194. Epub 2018 Mar 8.
本文引用的文献
1
The phosphoglucan phosphatase like sex Four2 dephosphorylates starch at the C3-position in Arabidopsis.磷酸葡聚糖磷酸酶样蛋白 SEX FOUR2 在拟南芥中于 C3 位去磷酸化淀粉。
Plant Cell. 2011 Nov;23(11):4096-111. doi: 10.1105/tpc.111.092155. Epub 2011 Nov 18.
2
Structural and functional analysis of PTPMT1, a phosphatase required for cardiolipin synthesis.PTPMT1 的结构与功能分析，一种合成心磷脂所必需的磷酸酶。
Proc Natl Acad Sci U S A. 2011 Jul 19;108(29):11860-5. doi: 10.1073/pnas.1109290108. Epub 2011 Jul 5.
3
The extended PP1 toolkit: designed to create specificity.扩展的 PP1 工具包：旨在提高特异性。
Trends Biochem Sci. 2010 Aug;35(8):450-8. doi: 10.1016/j.tibs.2010.03.002. Epub 2010 May 1.
4
Cracking the phosphatase code: docking interactions determine substrate specificity.破解磷酸酶密码：对接相互作用决定底物特异性。
Sci Signal. 2009 Dec 8;2(100):re9. doi: 10.1126/scisignal.2100re9.
5
Serine/threonine phosphatases: mechanism through structure.丝氨酸/苏氨酸磷酸酶：基于结构的作用机制
Cell. 2009 Oct 30;139(3):468-84. doi: 10.1016/j.cell.2009.10.006.
6
Conservation of the glucan phosphatase laforin is linked to rates of molecular evolution and the glucan metabolism of the organism.葡聚糖磷酸酶拉福林的保守性与分子进化速率及生物体的葡聚糖代谢相关。
BMC Evol Biol. 2009 Jun 22;9:138. doi: 10.1186/1471-2148-9-138.
7
From promiscuity to precision: protein phosphatases get a makeover.从杂乱无章到精准无误：蛋白磷酸酶焕然一新。
Mol Cell. 2009 Mar 13;33(5):537-45. doi: 10.1016/j.molcel.2009.02.015.
8
Dual-specificity phosphatases: critical regulators with diverse cellular targets.双特异性磷酸酶：具有多种细胞靶点的关键调节因子。
Biochem J. 2009 Mar 15;418(3):475-89. doi: 10.1042/bj20082234.
9
Evolution of protein phosphatases in plants and animals.植物和动物中蛋白质磷酸酶的进化
Biochem J. 2009 Jan 15;417(2):401-9. doi: 10.1042/BJ20081986.
10
Laforin is a glycogen phosphatase, deficiency of which leads to elevated phosphorylation of glycogen in vivo.拉福林是一种糖原磷酸酶，其缺乏会导致体内糖原磷酸化水平升高。
Proc Natl Acad Sci U S A. 2007 Dec 4;104(49):19262-6. doi: 10.1073/pnas.0707952104. Epub 2007 Nov 26.
Zotero 插件