Iterative in Situ Click Chemistry Assembles a Branched Capture Agent and Allosteric Inhibitor for Akt1
| Autor: | K. Barry Sharpless, Heather D. Agnew, Kaycie Deyle, James R. Heath, Jaehong Lim, Arundhati Nag, Su Seong Lee, Ryan K. Henning, Suresh M. Pitram, Steven W. Millward, Jessica A. Pfeilsticker, Gabriel A. Kwong, Jason E. Hein |
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| Rok vydání: | 2011 |
| Předmět: |
chemistry.chemical_classification
In situ Chemistry medicine.drug_class Allosteric regulation Peptide General Chemistry Ligand (biochemistry) Monoclonal antibody Biochemistry Small molecule Combinatorial chemistry Article Catalysis Structure-Activity Relationship Colloid and Surface Chemistry medicine Click chemistry Structure–activity relationship Click Chemistry Protein Kinase Inhibitors Proto-Oncogene Proteins c-akt Allosteric Site |
| Zdroj: | Journal of the American Chemical Society. 133:18280-18288 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/ja2064389 |
| Popis: | We describe the use of iterative in situ click chemistry to design an Akt-specific branched peptide triligand that is a drop-in replacement for monoclonal antibodies in multiple biochemical assays. Each peptide module in the branched structure makes unique contributions to affinity and/or specificity resulting in a 200 nM affinity ligand that efficiently immunoprecipitates Akt from cancer cell lysates and labels Akt in fixed cells. Our use of a small molecule to preinhibit Akt prior to screening resulted in low micromolar inhibitory potency and an allosteric mode of inhibition, which is evidenced through a series of competitive enzyme kinetic assays. To demonstrate the efficiency and selectivity of the protein-templated in situ click reaction, we developed a novel QPCR-based methodology that enabled a quantitative assessment of its yield. These results point to the potential for iterative in situ click chemistry to generate potent, synthetically accessible antibody replacements with novel inhibitory properties. |
| Databáze: | OpenAIRE |
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