ArticleACS chemical biology2009
Modular strategy for the semisynthesis of a K+ channel: investigating interactions of the pore helix.
Article in ACS chemical biology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 39 citations in OpenAlex.
- New Analogs of the Compstatin Family of Clinical Complement Inhibitors with Low Picomolar Target Affinity.Journal of medicinal chemistry · 2026Article
- "Lys-Leu" Motif Empowers ExpedientJACS Au · 2026Article
- Probing Ion Configurations in the KcsA Selectivity Filter with Single-Isotope Labels and 2D IR Spectroscopy.Journal of the American Chemical Society · 2023Article
- Incorporating thioamides into proteins by native chemical ligation.Methods in enzymology · 2021Article
- Expanding the Scope of Orthogonal Translation with Pyrrolysyl-tRNA Synthetases Dedicated to Aromatic Amino Acids.Molecules (Basel, Switzerland) · 2020Article
- Chemoenzymatic Semisynthesis of Proteins.Chemical reviews · 2020Review
- A Shared Mechanism for the Folding of Voltage-Gated KBiochemistry · 2019Article
- Probing the Effects of Gating on the Ion Occupancy of the KJournal of the American Chemical Society · 2017Article
- A semisynthetic Atg3 reveals that acetylation promotes Atg3 membrane binding and Atg8 lipidation.Nature communications · 2017Article
- Instantaneous ion configurations in the K+ ion channel selectivity filter revealed by 2D IR spectroscopy.Science (New York, N.Y.) · 2016Article
- Individual Ion Binding Sites in the K(+) Channel Play Distinct Roles in C-type Inactivation and in Recovery from Inactivation.Structure (London, England : 1993) · 2016Article
- Probing the active site tryptophan of Staphylococcus aureus thioredoxin with an analog.Nucleic acids research · 2015Article
- Semisynthetic protein nanoreactor for single-molecule chemistry.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Chemical ligation of the influenza M2 protein for solid-state NMR characterization of the cytoplasmic domain.Protein science : a publication of the Protein Society · 2015Article
- Histones: at the crossroads of peptide and protein chemistry.Chemical reviews · 2015Review
- Application of the protein semisynthesis strategy to the generation of modified chromatin.Annual review of biochemistry · 2015Review
- Hydrogels Formed by Oxo-ester Mediated Native Chemical Ligation.Biomaterials science · 2013Article
- Scorpion toxins specific for potassium (K+) channels: a historical overview of peptide bioengineering.Toxins · 2012Review
- Elucidating the role of C-terminal post-translational modifications using protein semisynthesis strategies: α-synuclein phosphorylation at tyrosine 125.Journal of the American Chemical Society · 2012Article
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Authors and funding
4 authors at 1 institution in 1 country.
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Abstract
Chemical synthesis is a powerful method for precise modification of the structural and electronic properties of proteins. The difficulties in the synthesis and purification of peptides containing transmembrane segments have presented obstacles to the chemical synthesis of integral membrane proteins. Here, we present a modular strategy for the semisynthesis of integral membrane proteins in which solid-phase peptide synthesis is limited to the region of interest, while the rest of the protein is obtained by recombinant means. This modular strategy considerably simplifies the synthesis and purification steps that have previously hindered the chemical synthesis of integral membrane proteins. We develop a SUMO fusion and proteolysis approach for obtaining the N-terminal cysteine containing membrane-spanning peptides required for the semisynthesis. We demonstrate the feasibility of the modular approach by the semisynthesis of full-length KcsA K(+) channels in which only regions of interest, such as the selectivity filter or the pore helix, are obtained by chemical synthesis. The modular approach is used to investigate the hydrogen bond interactions of a tryptophan residue in the pore helix, tryptophan 68, by substituting it with the isosteric analogue, beta-(3-benzothienyl)-l-alanine (3BT). A functional analysis of the 3BT mutant channels indicates that the K(+) conduction and selectivity of the 3BT mutant channels are similar to those of the wild type, but the mutant channels show a 3-fold increase in Rb(+) conduction. These results suggest that the hydrogen bond interactions of tryptophan 68 are essential for optimizing the selectivity filter for K(+) conduction over Rb(+) conduction.
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Registered trials
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.