一种用于鉴定蛋白酶体核心颗粒小分子刺激剂的灵敏高通量筛选方法。
A Sensitive High-Throughput Screening Method for Identifying Small Molecule Stimulators of the Core Particle of the Proteasome.
作者信息
Coleman Rachel A, Trader Darci J
机构信息
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, Indiana.
出版信息
Curr Protoc Chem Biol. 2018 Dec;10(4):e52. doi: 10.1002/cpch.52. Epub 2018 Oct 4.
Abstract
Fluorescence resonance energy transfer (FRET) technology is a useful tool to monitor protein interactions as well as protease activity. We have recently reported a biochemical assay utilizing a FRET reporter peptide to monitor the activity of the 20S catalytic particle (20S CP) of the proteasome. This assay is designed specifically to have increased sensitivity to identify stimulators of the 20S CP, which may hold therapeutic potential to treat protein-accumulation diseases. The protocol described here details the necessary steps in synthesizing the FRET reporter peptide and performing the FRET assay with the 20S CP. © 2018 by John Wiley & Sons, Inc.
摘要
荧光共振能量转移（FRET）技术是监测蛋白质相互作用以及蛋白酶活性的一种有用工具。我们最近报道了一种利用FRET报告肽监测蛋白酶体20S催化颗粒（20S CP）活性的生化检测方法。该检测方法经过专门设计，具有更高的灵敏度，可用于识别20S CP的刺激剂，这些刺激剂可能具有治疗蛋白质蓄积性疾病的潜在价值。此处描述的实验方案详细介绍了合成FRET报告肽以及用20S CP进行FRET检测的必要步骤。© 2018约翰威立国际出版公司
相似文献
1
A Sensitive High-Throughput Screening Method for Identifying Small Molecule Stimulators of the Core Particle of the Proteasome.
Curr Protoc Chem Biol. 2018 Dec;10(4):e52. doi: 10.1002/cpch.52. Epub 2018 Oct 4.
2
Development and Application of a Sensitive Peptide Reporter to Discover 20S Proteasome Stimulators.
ACS Comb Sci. 2018 May 14;20(5):269-276. doi: 10.1021/acscombsci.7b00193. Epub 2018 Mar 23.
3
Methods to Discover and Evaluate Proteasome Small Molecule Stimulators.
Molecules. 2019 Jun 25;24(12):2341. doi: 10.3390/molecules24122341.
4
Small Molecule Enhancement of 20S Proteasome Activity Targets Intrinsically Disordered Proteins.
ACS Chem Biol. 2017 Sep 15;12(9):2240-2247. doi: 10.1021/acschembio.7b00489. Epub 2017 Aug 1.
5
Approaches to Evaluate the Impact of a Small-Molecule Binder to a Noncatalytic Site of the Proteasome.
Chembiochem. 2021 Jun 2;22(11):1961-1965. doi: 10.1002/cbic.202100023. Epub 2021 Mar 26.
6
Small Molecule Modulation of Proteasome Assembly.
Biochemistry. 2018 Jul 17;57(28):4214-4224. doi: 10.1021/acs.biochem.8b00579. Epub 2018 Jun 27.
7
High-throughput amenable fluorescence-assays to screen for calmodulin-inhibitors.
Anal Biochem. 2019 May 1;572:25-32. doi: 10.1016/j.ab.2019.02.015. Epub 2019 Feb 27.
8
Establishment of a suite of assays that support the discovery of proteasome stimulators.
Biochim Biophys Acta Gen Subj. 2017 Apr;1861(4):892-899. doi: 10.1016/j.bbagen.2017.01.003. Epub 2017 Jan 5.
9
A FRET-based approach for identification of proteasome catalytic subunit composition.
Mol Biosyst. 2014 Feb;10(2):196-200. doi: 10.1039/c3mb70471h.
10
Small-Molecule Stimulators of NRF1 Transcriptional Activity.
Chembiochem. 2020 Jul 1;21(13):1816-1819. doi: 10.1002/cbic.201900487. Epub 2019 Nov 8.
引用本文的文献
1
Blm10-Based Compounds Add to the Knowledge of How Allosteric Modulators Influence Human 20S Proteasome.
ACS Chem Biol. 2025 Feb 21;20(2):266-280. doi: 10.1021/acschembio.4c00341. Epub 2025 Feb 5.
2
Primed for Interactions: Investigating the Primed Substrate Channel of the Proteasome for Improved Molecular Engagement.
Molecules. 2024 Jul 17;29(14):3356. doi: 10.3390/molecules29143356.
3
Development of a Cell-Based AlphaLISA Assay for High-Throughput Screening for Small Molecule Proteasome Modulators.
ACS Omega. 2023 Apr 21;8(17):15650-15659. doi: 10.1021/acsomega.3c01158. eCollection 2023 May 2.
4
Protein degradation profile reveals dynamic nature of 20S proteasome small molecule stimulation.
RSC Chem Biol. 2021 Jan 5;2(2):636-644. doi: 10.1039/d0cb00191k. eCollection 2021 Apr 1.
5
Proteasome Activation to Combat Proteotoxicity.
Molecules. 2019 Aug 5;24(15):2841. doi: 10.3390/molecules24152841.
本文引用的文献
1
Aim for the core: suitability of the ubiquitin-independent 20S proteasome as a drug target in neurodegeneration.
Transl Res. 2018 Aug;198:48-57. doi: 10.1016/j.trsl.2018.05.002. Epub 2018 Jun 19.
2
Development and Application of a Sensitive Peptide Reporter to Discover 20S Proteasome Stimulators.
ACS Comb Sci. 2018 May 14;20(5):269-276. doi: 10.1021/acscombsci.7b00193. Epub 2018 Mar 23.
3
Small Molecule Enhancement of 20S Proteasome Activity Targets Intrinsically Disordered Proteins.
ACS Chem Biol. 2017 Sep 15;12(9):2240-2247. doi: 10.1021/acschembio.7b00489. Epub 2017 Aug 1.
4
Establishment of a suite of assays that support the discovery of proteasome stimulators.
Biochim Biophys Acta Gen Subj. 2017 Apr;1861(4):892-899. doi: 10.1016/j.bbagen.2017.01.003. Epub 2017 Jan 5.
5
Regulating the 20S proteasome ubiquitin-independent degradation pathway.
Biomolecules. 2014 Sep 23;4(3):862-84. doi: 10.3390/biom4030862.
6
Regulated protein turnover: snapshots of the proteasome in action.
Nat Rev Mol Cell Biol. 2014 Feb;15(2):122-33. doi: 10.1038/nrm3741.
7
Visualization of human immunodeficiency virus protease inhibition using a novel Förster resonance energy transfer molecular probe.
Biotechnol Prog. 2011 Jul;27(4):1107-14. doi: 10.1002/btpr.628. Epub 2011 May 16.
8
Protein aggregation in the brain: the molecular basis for Alzheimer's and Parkinson's diseases.
Mol Med. 2008 Jul-Aug;14(7-8):451-64. doi: 10.2119/2007-00100.Irvine.
9
The ubiquitin-proteasome system.
J Biosci. 2006 Mar;31(1):137-55. doi: 10.1007/BF02705243.
10
Autophagy and related mechanisms of lysosome-mediated protein degradation.
Trends Cell Biol. 1994 Apr;4(4):139-43. doi: 10.1016/0962-8924(94)90069-8.
Zotero 插件