• 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

生成 Ca2+-独立型 sortase A 突变体，增强其对蛋白质和细胞表面标记的活性。

Generation of Ca2+-independent sortase A mutants with enhanced activity for protein and cell surface labeling.

机构信息

Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, Massachusetts, United States of America.

Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, United States of America.

出版信息

PLoS One. 2017 Dec 4;12(12):e0189068. doi: 10.1371/journal.pone.0189068. eCollection 2017.

DOI:10.1371/journal.pone.0189068
PMID:29200433
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5714338/
Abstract

Sortase A, a calcium-dependent transpeptidase derived from Staphylococcus aureus, is used in a broad range of applications, such as the conjugation of fluorescent dyes and other moieties to proteins or to the surface of eukaryotic cells. In vivo and cell-based applications of sortase have been somewhat limited by the large range of calcium concentrations, as well as by the often transient nature of protein-protein interactions in living systems. In order to use sortase A for cell labeling applications, we generated a new sortase A variant by combining multiple mutations to yield an enzyme that was both calcium-independent and highly active. This variant has enhanced activity for both N- and C-terminal labeling, as well as for cell surface modification under physiological conditions.

摘要

金黄色葡萄球菌来源的依赖钙的转肽酶 Sortase A 广泛应用于各种领域，例如将荧光染料和其他基团连接到蛋白质或真核细胞表面。Sortase 在体内和基于细胞的应用受到钙浓度范围广以及生物系统中蛋白质-蛋白质相互作用往往是短暂的限制。为了将 Sortase A 用于细胞标记应用，我们通过组合多个突变生成了一种新的 Sortase A 变体，该变体既不依赖钙又具有高度活性。该变体在生理条件下对 N 端和 C 端标记以及细胞表面修饰均具有增强的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/0ba53fd1f4a8/pone.0189068.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/b64ca441a4c3/pone.0189068.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/eb395ab297e7/pone.0189068.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/4f1dc4f772e1/pone.0189068.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/837dd08874af/pone.0189068.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/e50ac6dc9d0e/pone.0189068.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/28cce6225e72/pone.0189068.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/0ba53fd1f4a8/pone.0189068.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/b64ca441a4c3/pone.0189068.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/eb395ab297e7/pone.0189068.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/4f1dc4f772e1/pone.0189068.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/837dd08874af/pone.0189068.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/e50ac6dc9d0e/pone.0189068.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/28cce6225e72/pone.0189068.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4533/5714338/0ba53fd1f4a8/pone.0189068.g007.jpg

相似文献

1
Generation of Ca2+-independent sortase A mutants with enhanced activity for protein and cell surface labeling.生成 Ca2+-独立型 sortase A 突变体，增强其对蛋白质和细胞表面标记的活性。
PLoS One. 2017 Dec 4;12(12):e0189068. doi: 10.1371/journal.pone.0189068. eCollection 2017.
2
Design of Ca2+-independent Staphylococcus aureus sortase A mutants.设计不依赖 Ca2+ 的金黄色葡萄球菌 sortase A 突变体。
Biotechnol Bioeng. 2012 Dec;109(12):2955-61. doi: 10.1002/bit.24585. Epub 2012 Jul 4.
3
Site-specific protein modification using immobilized sortase in batch and continuous-flow systems.在分批和连续流动系统中使用固定化分选酶进行位点特异性蛋白质修饰。
Nat Protoc. 2015 Mar;10(3):508-16. doi: 10.1038/nprot.2015.026. Epub 2015 Feb 26.
4
Mutational analysis of active site residues in the Staphylococcus aureus transpeptidase SrtA.金黄色葡萄球菌转肽酶SrtA活性位点残基的突变分析
Biochemistry. 2007 Jun 19;46(24):7269-78. doi: 10.1021/bi700448e. Epub 2007 May 23.
5
Sortase mutants with improved protein thermostability and enzymatic activity obtained by consensus design.通过共有序列设计获得的具有改善的蛋白质热稳定性和酶活性的分选酶突变体。
Protein Eng Des Sel. 2019 Dec 31;32(12):555-564. doi: 10.1093/protein/gzaa018.
6
The Transpeptidase Sortase A Binds Nucleic Acids and Mediates Mammalian Cell Labeling.转肽酶 Sortase A 结合核酸并介导哺乳动物细胞标记。
Adv Sci (Weinh). 2024 Jun;11(21):e2305605. doi: 10.1002/advs.202305605. Epub 2024 Apr 5.
7
Anchoring foreign substances on live cell surfaces using Sortase A specific binding peptide.利用 Sortase A 特异性结合肽将外源物质锚定在活细胞表面。
Chem Commun (Camb). 2013 Oct 25;49(83):9585-7. doi: 10.1039/c3cc44753g.
8
Structure of sortase, the transpeptidase that anchors proteins to the cell wall of Staphylococcus aureus.分选酶的结构，这种转肽酶可将蛋白质锚定在金黄色葡萄球菌的细胞壁上。
Proc Natl Acad Sci U S A. 2001 May 22;98(11):6056-61. doi: 10.1073/pnas.101064198.
9
Mutagenesis studies of substrate recognition and catalysis in the sortase A transpeptidase from Staphylococcus aureus.金黄色葡萄球菌分选酶A转肽酶中底物识别与催化的诱变研究。
J Biol Chem. 2008 May 23;283(21):14762-71. doi: 10.1074/jbc.M800974200. Epub 2008 Mar 28.
10
Sortase: the surface protein anchoring transpeptidase and the LPXTG motif.分选酶：表面蛋白锚定转肽酶与LPXTG基序
Trends Microbiol. 2000 Apr;8(4):148-51. doi: 10.1016/s0966-842x(00)01741-8.

引用本文的文献

1
N‑Terminal Conjugation of a Near-Infrared Photosensitizer to an Antibody for Cancer Diagnosis and Treatment.用于癌症诊断与治疗的近红外光敏剂与抗体的N端共轭
ACS Omega. 2025 Jun 11;10(24):25596-25604. doi: 10.1021/acsomega.5c01083. eCollection 2025 Jun 24.
2
Recombinant-Chemosynthetic Biosensors for Probing Cell Surface Signaling of Red Blood Cells and Other Cells.用于探测红细胞和其他细胞表面信号传导的重组化学合成生物传感器
Chem Biomed Imaging. 2025 Jan 3;3(2):95-110. doi: 10.1021/cbmi.4c00067. eCollection 2025 Feb 24.
3
C-terminal amides mark proteins for degradation via SCF-FBXO31.

本文引用的文献

1
PD-L1 is an activation-independent marker of brown adipocytes.程序性死亡受体配体1（PD-L1）是棕色脂肪细胞的一种不依赖激活的标志物。
Nat Commun. 2017 Sep 21;8(1):647. doi: 10.1038/s41467-017-00799-8.
2
Site-Specific Protein Labeling via Sortase-Mediated Transpeptidation.通过分选酶介导的转肽作用进行位点特异性蛋白质标记。
Curr Protoc Protein Sci. 2017 Aug 1;89:15.3.1-15.3.19. doi: 10.1002/cpps.38.
3
Engineered erythrocytes covalently linked to antigenic peptides can protect against autoimmune disease.与抗原肽共价连接的工程红细胞可预防自身免疫性疾病。
C末端酰胺化标记蛋白质以便通过SCF-FBXO31进行降解。
Nature. 2025 Feb;638(8050):519-527. doi: 10.1038/s41586-024-08475-w. Epub 2025 Jan 29.
4
CD45-PET is a robust, non-invasive tool for imaging inflammation.CD45正电子发射断层扫描是一种用于炎症成像的强大的非侵入性工具。
Nature. 2025 Mar;639(8053):214-224. doi: 10.1038/s41586-024-08441-6. Epub 2025 Jan 22.
5
Site-Specific and Fluorescently Enhanced Installation of Post-Translational Protein Modifications via Bifunctional Biarsenical Linker.通过双功能双砷连接子进行翻译后蛋白质修饰的位点特异性和荧光增强安装
ACS Omega. 2024 Oct 30;9(45):45127-45137. doi: 10.1021/acsomega.4c05828. eCollection 2024 Nov 12.
6
Versatile tissue-injectable hydrogels capable of the extended hydrolytic release of bioactive protein therapeutics.能够实现生物活性蛋白治疗药物延长水解释放的多功能可注射组织水凝胶。
Bioeng Transl Med. 2024 Apr 15;9(5):e10668. doi: 10.1002/btm2.10668. eCollection 2024 Sep.
7
Regulation of minimal spindle midzone organization by mitotic kinases.有丝分裂激酶对最小纺锤体中部组织的调控。
Nat Commun. 2024 Oct 29;15(1):9213. doi: 10.1038/s41467-024-53500-1.
8
Alkylamine-tethered molecules recruit FBXO22 for targeted protein degradation.烷胺连接的分子招募 FBXO22 进行靶向蛋白降解。
Nat Commun. 2024 Jun 26;15(1):5409. doi: 10.1038/s41467-024-49739-3.
9
A monoclonal antibody that recognizes a unique 13-residue epitope in the cytoplasmic tail of HLA-E.一种单克隆抗体，可识别HLA-E细胞质尾部一个独特的13个氨基酸残基的表位。
Mol Immunol. 2024 Aug;172:56-67. doi: 10.1016/j.molimm.2024.06.004. Epub 2024 Jun 19.
10
Nanobody-based CAR NK cells for possible immunotherapy of MICA tumors.基于纳米抗体的嵌合抗原受体自然杀伤细胞用于MICA肿瘤的潜在免疫治疗。
PNAS Nexus. 2024 May 6;3(5):pgae184. doi: 10.1093/pnasnexus/pgae184. eCollection 2024 May.
Proc Natl Acad Sci U S A. 2017 Mar 21;114(12):3157-3162. doi: 10.1073/pnas.1701746114. Epub 2017 Mar 7.
4
Hepta-Mutant Staphylococcus aureus Sortase A (SrtA) as a Tool for in Vivo Protein Labeling in Caenorhabditis elegans.七突变体金黄色葡萄球菌分选酶A（SrtA）作为秀丽隐杆线虫体内蛋白质标记的工具。
ACS Chem Biol. 2017 Mar 17;12(3):664-673. doi: 10.1021/acschembio.6b00998. Epub 2017 Jan 18.
5
Butelase-mediated cyclization and ligation of peptides and proteins.Butelase介导的肽和蛋白质的环化与连接。
Nat Protoc. 2016 Oct;11(10):1977-1988. doi: 10.1038/nprot.2016.118. Epub 2016 Sep 22.
6
Improved variants of SrtA for site-specific conjugation on antibodies and proteins with high efficiency.高效定点偶联抗体和蛋白质的 SrtA 改良变体。
Sci Rep. 2016 Aug 18;6:31899. doi: 10.1038/srep31899.
7
Recent advances in sortase-catalyzed ligation methodology.分选酶催化连接方法的最新进展。
Curr Opin Struct Biol. 2016 Jun;38:111-8. doi: 10.1016/j.sbi.2016.05.021. Epub 2016 Jun 16.
8
In Vivo Site-Specific Protein Tagging with Diverse Amines Using an Engineered Sortase Variant.利用工程化的 sortase 变体对活体中具有不同胺的特异性蛋白质进行标记。
J Am Chem Soc. 2016 Jun 22;138(24):7496-9. doi: 10.1021/jacs.6b03836. Epub 2016 Jun 9.
9
The use of F-2-fluorodeoxyglucose (FDG) to label antibody fragments for immuno-PET of pancreatic cancer.使用F-2-氟脱氧葡萄糖（FDG）标记抗体片段用于胰腺癌的免疫正电子发射断层显像（immuno-PET）。
ACS Cent Sci. 2015 Jun 24;1(3):142-147. doi: 10.1021/acscentsci.5b00121. Epub 2015 Jun 3.
10
Ca2+ -independent sortase-A exhibits high selective protein ligation activity in the cytoplasm of Escherichia coli.不依赖Ca2+的分选酶A在大肠杆菌细胞质中表现出高选择性蛋白质连接活性。
Biotechnol J. 2015 Sep;10(9):1487-92. doi: 10.1002/biot.201500012. Epub 2015 Apr 28.