ArticleJournal of structural and functional genomics2010
Cleavable C-terminal His-tag vectors for structure determination.
Article in Journal of structural and functional genomics, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- New Insights into Binding of G-segment DNA to the Active Site of Escherichia coli Topoisomerase III.Scientific reports · 2026Article
- Expression, purification, and characterisation of the p53 binding domain of Retinoblastoma binding protein 6 (RBBP6).PloS one · 2023Article
- The use of high-affinity polyhistidine binders as masking probes for the selection of an NDM-1 specific aptamer.Scientific reports · 2022Article
- PABP1 Drives the Selective Translation of Influenza A Virus mRNA.Journal of molecular biology · 2022Article
- Crystal structure of betaine aldehyde dehydrogenase from Burkholderia pseudomallei.Acta crystallographica. Section F, Structural biology communications · 2022Article
- Article
- Influenza A Virus NS1 Protein Binds as a Dimer to RNA-Free PABP1 but Not to the PABP1·Poly(A) RNA Complex.Biochemistry · 2020Article
- 3D domain swapping in the TIM barrel of the α subunit of Streptococcus pneumoniae tryptophan synthase.Acta crystallographica. Section D, Structural biology · 2020Article
- Structures of teixobactin-producing nonribosomal peptide synthetase condensation and adenylation domains.Current research in structural biology · 2020Article
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- Article
- Functional plasticity of antibacterial EndoU toxins.Molecular microbiology · 2018Article
- RNA Modulates the Interaction between Influenza A Virus NS1 and Human PABP1.Biochemistry · 2018Article
- Catalytic and structural properties of pheophytinase, the phytol esterase involved in chlorophyll breakdown.Journal of experimental botany · 2018Article
- PKS-NRPS Enzymology and Structural Biology: Considerations in Protein Production.Methods in enzymology · 2018Article
- Structure of a novel antibacterial toxin that exploits elongation factor Tu to cleave specific transfer RNAs.Nucleic acids research · 2017Article
- New ligation independent cloning vectors for expression of recombinant proteins with a self-cleaving CPD/6xHis-tag.BMC biotechnology · 2017Article
- Cellularly active N-hydroxyurea FEN1 inhibitors block substrate entry to the active site.Nature chemical biology · 2016Article
- High Level Expression and Purification of Recombinant Proteins from Escherichia coli with AK-TAG.PloS one · 2016Article
- A Chimeric Affinity Tag for Efficient Expression and Chromatographic Purification of Heterologous Proteins from Plants.Frontiers in plant science · 2016Article
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
High-throughput structural genomics projects seek to delineate protein structure space by determining the structure of representatives of all major protein families. Generally this is accomplished by processing numerous proteins through standardized protocols, for the most part involving purification of N-terminally His-tagged proteins. Often proteins that fail this approach are abandoned, but in many cases further effort is warranted because of a protein's intrinsic value. In addition, failure often occurs relatively far into the path to structure determination, and many failed proteins passed the first critical step, expression as a soluble protein. Salvage pathways seek to recoup the investment in this subset of failed proteins through alternative cloning, nested truncations, chemical modification, mutagenesis, screening buffers, ligands and modifying processing steps. To this end we have developed a series of ligation-independent cloning expression vectors that append various cleavable C-terminal tags instead of the conventional N-terminal tags. In an initial set of 16 proteins that failed with an N-terminal appendage, structures were obtained for C-terminally tagged derivatives of five proteins, including an example for which several alternative salvaging steps had failed. The new vectors allow appending C-terminal His(6)-tag and His(6)- and MBP-tags, and are cleavable with TEV or with both TEV and TVMV proteases.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.