• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

N 端多聚组氨酸标签对蛋白质热稳定性的影响。

Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability.

作者信息

Booth William T, Schlachter Caleb R, Pote Swanandi, Ussin Nikita, Mank Nicholas J, Klapper Vincent, Offermann Lesa R, Tang Chuanbing, Hurlburt Barry K, Chruszcz Maksymilian

机构信息

Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208, United States.

Department of Chemistry, Davidson College, Davidson, North Carolina 28035, United States.

出版信息

ACS Omega. 2018 Jan 31;3(1):760-768. doi: 10.1021/acsomega.7b01598. Epub 2018 Jan 22.

DOI:10.1021/acsomega.7b01598
PMID:29399652
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5793033/
Abstract

For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability.

摘要

多年来，多组氨酸标签（His标签）的使用一直是固定化金属亲和色谱实验中分离重组蛋白的主要方法。它们的使用在提高粗细胞裂解物中蛋白质纯度方面具有广泛的益处。对于一些重组蛋白来说，添加His标签的一个后果是它会影响蛋白质的功能和稳定性。功能性蛋白质对于阐明其生物学、动力学、结构和热力学性质至关重要。在本研究中，我们使用差示扫描荧光法测定了N端His标签对选定蛋白质热稳定性的影响，并发现去除His标签对其稳定性可能既有有益影响也有有害影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/897b4f54ce81/ao-2017-01598a_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/27099dfaa325/ao-2017-01598a_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/2b2ab69f4773/ao-2017-01598a_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/f212434f4138/ao-2017-01598a_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/897b4f54ce81/ao-2017-01598a_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/27099dfaa325/ao-2017-01598a_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/2b2ab69f4773/ao-2017-01598a_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/f212434f4138/ao-2017-01598a_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f77/6645752/897b4f54ce81/ao-2017-01598a_0003.jpg

相似文献

1
Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability.N 端多聚组氨酸标签对蛋白质热稳定性的影响。
ACS Omega. 2018 Jan 31;3(1):760-768. doi: 10.1021/acsomega.7b01598. Epub 2018 Jan 22.
2
Removal of poly-histidine fusion tags from recombinant proteins purified by expanded bed adsorption.从通过扩张床吸附纯化的重组蛋白中去除多组氨酸融合标签。
Biotechnol Bioeng. 2005 Nov 20;92(4):501-13. doi: 10.1002/bit.20633.
3
Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119.N端和C端多聚组氨酸标签对嗜热细胞色素P450 CYP119稳定性和功能的影响
Bioinorg Chem Appl. 2019 Jun 20;2019:8080697. doi: 10.1155/2019/8080697. eCollection 2019.
4
A new metal affinity NCTR tag as a better alternative to the His-tag for the expression of recombinant fused proteins.一种新型金属亲和NCTR标签，作为His标签的更好替代物用于重组融合蛋白的表达。
Protein Expr Purif. 2019 Dec;164:105477. doi: 10.1016/j.pep.2019.105477. Epub 2019 Aug 13.
5
Structural and biochemical insights into His-tag-induced higher-order oligomerization of membrane proteins by cryo-EM and size exclusion chromatography.通过冷冻电镜和尺寸排阻色谱对His标签诱导的膜蛋白高阶寡聚化的结构和生化见解。
J Struct Biol. 2023 Mar;215(1):107924. doi: 10.1016/j.jsb.2022.107924. Epub 2022 Nov 30.
6
Purification of a Recombinant Polyhistidine-Tagged Glucosyltransferase Using Immobilized Metal-Affinity Chromatography (IMAC).使用固定化金属亲和色谱法（IMAC）纯化重组多组氨酸标签葡萄糖基转移酶
Methods Mol Biol. 2016;1405:91-7. doi: 10.1007/978-1-4939-3393-8_9.
7
Effects of IMAC specific peptide tags on the stability of recombinant green fluorescent protein.IMAC 特异性肽标签对重组绿色荧光蛋白稳定性的影响。
Biotechnol Prog. 2011 Jul;27(4):1048-53. doi: 10.1002/btpr.546. Epub 2011 Jun 3.
8
The Effect of Recombinant Tags on Flavonol-Specific 3-O Glucosyltransferase Activity.重组标签对黄酮醇特异性3-O-葡萄糖基转移酶活性的影响。
Plants (Basel). 2020 Mar 24;9(3):402. doi: 10.3390/plants9030402.
9
Rapid purification of the potato ADP-glucose pyrophosphorylase by polyhistidine-mediated chromatography.通过多组氨酸介导的色谱法快速纯化马铃薯 ADP - 葡萄糖焦磷酸化酶。
Protein Expr Purif. 2004 Nov;38(1):99-107. doi: 10.1016/j.pep.2004.07.018.
10
Purification of Polyhistidine-Tagged Proteins.多组氨酸标签蛋白的纯化
Methods Mol Biol. 2017;1485:275-303. doi: 10.1007/978-1-4939-6412-3_14.

引用本文的文献

1
Expression and Purification of Recombinant Respiratory Syncytial Virus Proteins.重组呼吸道合胞病毒蛋白的表达与纯化
Methods Mol Biol. 2025;2948:137-157. doi: 10.1007/978-1-0716-4666-3_10.
2
Facile expression of proteins with desired N-terminal amino acid via an engineered cysteine protease domain.通过工程化的半胱氨酸蛋白酶结构域轻松表达具有所需N端氨基酸的蛋白质。
Commun Biol. 2025 Aug 6;8(1):1165. doi: 10.1038/s42003-025-08614-7.
3
C-terminal tagging impairs AGO2 function.C 端标记会损害AGO2的功能。

本文引用的文献

1
Streptococcus pyogenes quinolinate-salvage pathway-structural and functional studies of quinolinate phosphoribosyl transferase and NH -dependent NAD synthetase.化脓性链球菌喹啉酸挽救途径——喹啉酸磷酸核糖基转移酶和NH依赖性NAD合成酶的结构与功能研究
FEBS J. 2017 Aug;284(15):2425-2441. doi: 10.1111/febs.14136. Epub 2017 Jul 7.
2
Structural, Functional, and Immunological Characterization of Profilin Panallergens Amb a 8, Art v 4, and Bet v 2.肌动蛋白结合蛋白泛过敏原Amb a 8、Art v 4和Bet v 2的结构、功能及免疫学特征
J Biol Chem. 2016 Jul 22;291(30):15447-59. doi: 10.1074/jbc.M116.733659. Epub 2016 May 26.
3
Protein stability: a crystallographer's perspective.
RNA Biol. 2025 Dec;22(1):1-24. doi: 10.1080/15476286.2025.2534028. Epub 2025 Jul 23.
4
Immobilization of Thrombin on Agarose-Based Supports for Affinity Tag Removal.将凝血酶固定在基于琼脂糖的载体上用于去除亲和标签
Biomacromolecules. 2025 Jul 14;26(7):4718-4729. doi: 10.1021/acs.biomac.5c01010. Epub 2025 Jul 1.
5
Establishment of an efficient one-step enzymatic synthesis of cyclic-2,3-diphosphoglycerate.高效一步酶促合成环2,3-二磷酸甘油酸的方法建立。
Front Microbiol. 2025 May 21;16:1601972. doi: 10.3389/fmicb.2025.1601972. eCollection 2025.
6
IFIT2-induced transcriptomic changes in infected macrophages.IFIT2诱导感染巨噬细胞中的转录组变化。
Front Cell Infect Microbiol. 2025 May 20;15:1536446. doi: 10.3389/fcimb.2025.1536446. eCollection 2025.
7
A distinct class of ferredoxin:NADP oxidoreductase enzymes driving thermophilic ethanol production.一类独特的铁氧化还原蛋白：NADP氧化还原酶驱动嗜热乙醇生产。
J Biol Chem. 2025 May 21;301(7):110263. doi: 10.1016/j.jbc.2025.110263.
8
Yeast Complementation Assays Demonstrating the Importance of the Affinity Tag Position in Membrane Protein Purification, as Exemplified by UreI, the pH-Gated Urea Channel of .酵母互补分析证明了亲和标签位置在膜蛋白纯化中的重要性，以嗜热栖热菌的pH门控尿素通道UreI为例。
Small Sci. 2025 Jan 27;5(5):2400571. doi: 10.1002/smsc.202400571. eCollection 2025 May.
9
Bacterial suppressor-of-copper-sensitivity proteins exhibit diverse thiol-disulfide oxidoreductase cellular functions.细菌铜敏感性抑制蛋白具有多种硫醇-二硫键氧化还原酶细胞功能。
iScience. 2024 Nov 15;27(12):111392. doi: 10.1016/j.isci.2024.111392. eCollection 2024 Dec 20.
10
Comparative studies of seafood and reptile α- and β-parvalbumins.海鲜和爬行动物 α-和 β-副肌球蛋白的比较研究。
Protein Sci. 2024 Dec;33(12):e5226. doi: 10.1002/pro.5226.
蛋白质稳定性：晶体学家的视角
Acta Crystallogr F Struct Biol Commun. 2016 Feb;72(Pt 2):72-95. doi: 10.1107/S2053230X15024619. Epub 2016 Jan 26.
4
DSF Guided Refolding As A Novel Method Of Protein Production.二硫苏糖醇引导的重折叠作为一种新型蛋白质生产方法
Sci Rep. 2016 Jan 19;6:18906. doi: 10.1038/srep18906.
5
Protein purification and crystallization artifacts: The tale usually not told.蛋白质纯化和结晶假象：通常不为人知的故事。
Protein Sci. 2016 Mar;25(3):720-33. doi: 10.1002/pro.2861. Epub 2016 Jan 26.
6
Structural and Functional Characterization of the Hazelnut Allergen Cor a 8.榛子过敏原Cor a 8的结构与功能特性
J Agric Food Chem. 2015 Oct 21;63(41):9150-8. doi: 10.1021/acs.jafc.5b03534. Epub 2015 Oct 7.
7
Elusive Structural, Functional, and Immunological Features of Act d 5, the Green Kiwifruit Kiwellin.Act d 5，即绿色奇异果 kiwellin 的难以捉摸的结构、功能和免疫学特征。
J Agric Food Chem. 2015 Jul 29;63(29):6567-76. doi: 10.1021/acs.jafc.5b02159. Epub 2015 Jul 21.
8
Double trouble-Buffer selection and His-tag presence may be responsible for nonreproducibility of biomedical experiments.双重麻烦——缓冲液选择和组氨酸标签的存在可能导致生物医学实验的不可重复性。
Protein Sci. 2014 Oct;23(10):1359-68. doi: 10.1002/pro.2520. Epub 2014 Jul 30.
9
Structure and function of the peanut panallergen Ara h 8.花生过敏原 Ara h 8 的结构与功能。
J Biol Chem. 2013 Dec 27;288(52):36890-901. doi: 10.1074/jbc.M113.517797. Epub 2013 Nov 19.
10
Detrimental effect of the 6 His C-terminal tag on YedY enzymatic activity and influence of the TAT signal sequence on YedY synthesis.6 His C 末端标签对 YedY 酶活性的有害影响和 TAT 信号序列对 YedY 合成的影响。
BMC Biochem. 2013 Nov 1;14:28. doi: 10.1186/1471-2091-14-28.