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胞质磷脂酶Aα C2结构域以可溶形式表达后的纯化 。 你提供的原文似乎不完整，最后的“in.”后面应该还有具体内容。

Purification of Cytosolic Phospholipase Aα C2-domain after Expression in Soluble Form in .

作者信息

Hirano Yoshinori, Gao Yong-Guang, K Simanshu Dhirendra, J Stephenson Daniel, T Vu Ngoc, Malinina Lucy, E Chalfant Charles, J Patel Dinshaw, E Brown Rhoderick

机构信息

Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, U.S.A.

Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), Takayama, Japan.

出版信息

Bio Protoc. 2021 Feb 5;11(3):e3906. doi: 10.21769/BioProtoc.3906.

DOI:10.21769/BioProtoc.3906
PMID:33732793
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7952920/
Abstract

Previous expression/purification strategies for cytosolic phospholipase Aα C2-domain in have relied on refolded protein recovered from inclusion bodies and sometimes containing C-terminal Cys139Ala and Cys141Ser substitutions to eliminate potential refolding complications induced by Cys residues. The protocol presented herein describes an effective method for the expression of cytosolic phospholipase Aα C2-domain in soluble form in and subsequent purification to homogeneity. This protocol, which utilizes a cleavable 6xHis-SUMO tag, has recently been used to gain insights into the structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase Aα ( Hirano , 2019 ).

摘要

以往在大肠杆菌中表达/纯化胞质磷脂酶Aα C2结构域的策略依赖于从包涵体中回收的重折叠蛋白，该蛋白有时含有C端Cys139Ala和Cys141Ser取代，以消除由半胱氨酸残基引起的潜在重折叠复杂性。本文介绍的方案描述了一种在大肠杆菌中以可溶形式表达胞质磷脂酶Aα C2结构域并随后纯化至均一性的有效方法。该方案利用了一种可切割的6xHis-SUMO标签，最近已被用于深入了解胞质磷脂酶Aα的C2结构域识别磷脂酰胆碱的结构基础（Hirano，2019）。

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本文引用的文献

1
Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase Aα.胞质型磷脂酶 A2α的 C2 结构域识别磷脂酰胆碱的结构基础。
Elife. 2019 May 3;8:e44760. doi: 10.7554/eLife.44760.
2
Phospholipase A catalysis and lipid mediator lipidomics.磷脂酶 A 催化作用与脂质介质脂质组学。
Biochim Biophys Acta Mol Cell Biol Lipids. 2019 Jun;1864(6):766-771. doi: 10.1016/j.bbalip.2018.08.010. Epub 2018 Aug 23.
3
Selectivity of phospholipid hydrolysis by phospholipase A enzymes in activated cells leading to polyunsaturated fatty acid mobilization.激活细胞中磷脂酶 A 酶对磷脂的水解选择性导致多不饱和脂肪酸的动员。
Biochim Biophys Acta Mol Cell Biol Lipids. 2019 Jun;1864(6):772-783. doi: 10.1016/j.bbalip.2018.07.002. Epub 2018 Jul 17.
4
Review of four major distinct types of human phospholipase A.四种主要不同类型的人类磷脂酶A综述。
Adv Biol Regul. 2018 Jan;67:212-218. doi: 10.1016/j.jbior.2017.10.009. Epub 2017 Oct 23.
5
Cytosolic phospholipase A₂: physiological function and role in disease.胞质型磷脂酶A₂：生理功能及在疾病中的作用
J Lipid Res. 2015 Aug;56(8):1386-402. doi: 10.1194/jlr.R057588. Epub 2015 Apr 2.
6
Membranes serve as allosteric activators of phospholipase A2, enabling it to extract, bind, and hydrolyze phospholipid substrates.膜作为磷脂酶A2的变构激活剂，使其能够提取、结合并水解磷脂底物。
Proc Natl Acad Sci U S A. 2015 Feb 10;112(6):E516-25. doi: 10.1073/pnas.1424651112. Epub 2015 Jan 26.
7
The molecular basis of ceramide-1-phosphate recognition by C2 domains.鞘氨醇-1-磷酸被 C2 结构域识别的分子基础。
J Lipid Res. 2013 Mar;54(3):636-648. doi: 10.1194/jlr.M031088. Epub 2012 Dec 31.
8
Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention.磷脂酶A2 酶：物理结构、生物学功能、疾病关联、化学抑制及治疗干预
Chem Rev. 2011 Oct 12;111(10):6130-85. doi: 10.1021/cr200085w. Epub 2011 Sep 12.
9
SUMO fusion technology for enhanced protein expression and purification in prokaryotes and eukaryotes.用于增强原核生物和真核生物中蛋白质表达与纯化的SUMO融合技术。
Methods Mol Biol. 2011;705:15-30. doi: 10.1007/978-1-61737-967-3_2.
10
Localization and function of cytosolic phospholipase A2alpha at the Golgi.细胞质型磷脂酶 A2alpha 在高尔基体内的定位和功能。
Biochimie. 2010 Jun;92(6):620-6. doi: 10.1016/j.biochi.2010.03.001. Epub 2010 Mar 10.

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