一种用于简便纯化和单分子固定的肽标签系统。
A peptide tag system for facile purification and single-molecule immobilization.
机构信息
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637, USA.
出版信息
Biochemistry. 2009 Dec 22;48(50):11834-6. doi: 10.1021/bi901756n.
Abstract
A peptide fusion tag and accompanying recombinant capture reagents have been developed on the basis of the peptide-PDZ domain interaction and affinity clamps, a new class of affinity reagent. This system allows for single-step purification under mild conditions and stable capture of a tagged protein. The subnanomolar affinity, high force resistance (>30 pN), small size ( approximately 25 kDa, approximately one-sixth of the size of IgG), and monomeric nature of the affinity clamp are all superior features for many applications, in particular single-molecule measurements.
摘要
基于肽-PDZ 结构域相互作用和亲和夹，一种新型的亲和试剂，开发了一种肽融合标签和配套的重组捕获试剂。该系统允许在温和条件下进行单步纯化，并稳定捕获标记蛋白。亲和夹具有亚纳摩尔亲和力、高阻力 (>30 pN)、小尺寸（约 25 kDa，约为 IgG 大小的六分之一）和单体性质，这些都是许多应用的优异特性，特别是单分子测量。
相似文献
1
A peptide tag system for facile purification and single-molecule immobilization.
Biochemistry. 2009 Dec 22;48(50):11834-6. doi: 10.1021/bi901756n.
2
Development of new biotin/streptavidin reagents for pretargeting.
Biomol Eng. 1999 Dec 31;16(1-4):113-8. doi: 10.1016/s1050-3862(99)00044-3.
3
Mutagenesis of a flexible loop in streptavidin leads to higher affinity for the Strep-tag II peptide and improved performance in recombinant protein purification.
Protein Eng. 1997 Aug;10(8):975-82. doi: 10.1093/protein/10.8.975.
4
The Role of Changing Loop Conformations in Streptavidin Versions Engineered for High-affinity Binding of the Strep-tag II Peptide.
J Mol Biol. 2021 Apr 30;433(9):166893. doi: 10.1016/j.jmb.2021.166893. Epub 2021 Feb 24.
5
Stable, high-affinity streptavidin monomer for protein labeling and monovalent biotin detection.
Biotechnol Bioeng. 2013 Jan;110(1):57-67. doi: 10.1002/bit.24605. Epub 2012 Aug 8.
6
One-step purification of recombinant proteins using a nanomolar-affinity streptavidin-binding peptide, the SBP-Tag.
Protein Expr Purif. 2001 Dec;23(3):440-6. doi: 10.1006/prep.2001.1515.
7
Engineering Streptavidin and a Streptavidin-Binding Peptide with Infinite Binding Affinity and Reversible Binding Capability: Purification of a Tagged Recombinant Protein to High Purity via Affinity-Driven Thiol Coupling.
PLoS One. 2015 Sep 25;10(9):e0139137. doi: 10.1371/journal.pone.0139137. eCollection 2015.
8
Applications of a peptide ligand for streptavidin: the Strep-tag.
Biomol Eng. 1999 Dec 31;16(1-4):79-86. doi: 10.1016/s1050-3862(99)00033-9.
9
The random peptide library-assisted engineering of a C-terminal affinity peptide, useful for the detection and purification of a functional Ig Fv fragment.
Protein Eng. 1993 Jan;6(1):109-22. doi: 10.1093/protein/6.1.109.
10
Genetic engineering of streptavidin, a versatile affinity tag.
J Chromatogr B Biomed Sci Appl. 1998 Sep 11;715(1):85-91. doi: 10.1016/s0378-4347(98)00316-8.
引用本文的文献
1
BioID in Bacteria: Selection of a Suitable Biotin Ligase for Proxeome Mapping.
Methods Mol Biol. 2025;2953:17-39. doi: 10.1007/978-1-0716-4694-6_2.
2
Discovery of Therapeutic Antibodies Targeting Complex Multi-Spanning Membrane Proteins.
BioDrugs. 2024 Nov;38(6):769-794. doi: 10.1007/s40259-024-00682-1. Epub 2024 Oct 25.
3
Purification and Quality Control of Recombinant Proteins Expressed in Mammalian Cells: A Practical Review.
Methods Mol Biol. 2024;2810:329-353. doi: 10.1007/978-1-0716-3878-1_21.
4
Homologous mutations in human β, embryonic, and perinatal muscle myosins have divergent effects on molecular power generation.
Proc Natl Acad Sci U S A. 2024 Feb 27;121(9):e2315472121. doi: 10.1073/pnas.2315472121. Epub 2024 Feb 20.
5
Motility Assay to Probe the Calcium Sensitivity of Myosin and Regulated Thin Filaments.
Methods Mol Biol. 2024;2735:169-189. doi: 10.1007/978-1-0716-3527-8_10.
6
Trivalent SARS-CoV-2 S1 Subunit Protein Vaccination Induces Broad Humoral Responses in BALB/c Mice.
Vaccines (Basel). 2023 Jan 31;11(2):314. doi: 10.3390/vaccines11020314.
7
Adenovirus-vectored SARS-CoV-2 vaccine expressing S1-N fusion protein.
Antib Ther. 2022 Jul 8;5(3):177-191. doi: 10.1093/abt/tbac015. eCollection 2022 Jul.
8
Small Peptide-Protein Interaction Pair for Genetically Encoded, Fixation Compatible Peptide-PAINT.
Nano Lett. 2021 Nov 24;21(22):9509-9516. doi: 10.1021/acs.nanolett.1c02895. Epub 2021 Nov 10.
9
Identification of sequence changes in myosin II that adjust muscle contraction velocity.
PLoS Biol. 2021 Jun 10;19(6):e3001248. doi: 10.1371/journal.pbio.3001248. eCollection 2021 Jun.
10
Myosin motor domains carrying mutations implicated in early or late onset hypertrophic cardiomyopathy have similar properties.
J Biol Chem. 2019 Nov 15;294(46):17451-17462. doi: 10.1074/jbc.RA119.010563. Epub 2019 Oct 3.
本文引用的文献
1
Structural basis for exquisite specificity of affinity clamps, synthetic binding proteins generated through directed domain-interface evolution.
J Mol Biol. 2009 Oct 9;392(5):1221-31. doi: 10.1016/j.jmb.2009.07.067. Epub 2009 Jul 30.
2
A myosin motor that selects bundled actin for motility.
Proc Natl Acad Sci U S A. 2008 Jul 15;105(28):9616-20. doi: 10.1073/pnas.0802592105. Epub 2008 Jul 3.
3
The structure of the anti-c-myc antibody 9E10 Fab fragment/epitope peptide complex reveals a novel binding mode dominated by the heavy chain hypervariable loops.
Proteins. 2008 Nov 15;73(3):552-65. doi: 10.1002/prot.22080.
4
Design of protein function leaps by directed domain interface evolution.
Proc Natl Acad Sci U S A. 2008 May 6;105(18):6578-83. doi: 10.1073/pnas.0801097105. Epub 2008 Apr 29.
5
Single-molecule DNA nanomanipulation: improved resolution through use of shorter DNA fragments.
Nat Methods. 2005 Feb;2(2):127-38. doi: 10.1038/nmeth0205-127.
6
Making the most of affinity tags.
Trends Biotechnol. 2005 Jun;23(6):316-20. doi: 10.1016/j.tibtech.2005.03.012.
7
Origins of PDZ domain ligand specificity. Structure determination and mutagenesis of the Erbin PDZ domain.
J Biol Chem. 2003 Feb 28;278(9):7645-54. doi: 10.1074/jbc.M209751200. Epub 2002 Nov 20.
8
Myosin VI is a processive motor with a large step size.
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13655-9. doi: 10.1073/pnas.191512398. Epub 2001 Nov 13.
9
Affinity of the monoclonal antibody M1 directed against the FLAG peptide.
J Chromatogr A. 2001 Jun 29;921(1):25-30. doi: 10.1016/s0021-9673(01)00831-7.
10
Myosin-X, a novel myosin with pleckstrin homology domains, associates with regions of dynamic actin.
J Cell Sci. 2000 Oct;113 Pt 19:3439-51. doi: 10.1242/jcs.113.19.3439.
Zotero 插件