an-vitro-enzymatic-assay-to-measure-transcription-inhibition-gallium

An <em>In Vitro</em> Enzymatic Assay to Measure Transcription Inhibition by Gallium(III) and H<sub>3</sub> 5,10,15-tris(pentafluorophenyl)corroles

Chemotherapy often involves broad-spectrum cytotoxic agents with many side effects and limited targeting. Corroles are a class of tetrapyrrolic ...

California Institute of Technology

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.

Ryan K. Henning

Division of Chemistry and Chemical Engineering

California Institute of Technology

An In Vitro Enzymatic Assay to Measure Transcription Inhibition by Gallium(III) and H₃ 5,10,15-tris(pentafluorophenyl)corroles

Ryan K. Henning

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Division of Chemistry and Chemical Engineering

California Institute of Technology

An In Vitro Enzymatic Assay to Measure Transcription Inhibition by Gallium(III) and H3 5,10,15-tris(pentafluorophenyl)corroles

Division of Chemistry and Chemical Engineering

An In Vitro Enzymatic Assay to Measure Transcription Inhibition by Gallium(III) and H₃ 5,10,15-tris(pentafluorophenyl)corroles