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Sensing Enzymatic Activity by Exposure and Selection of DNA-Encoded Probes.

ABSTRACT: A sensing approach is applied to encode quantitative enzymatic activity information into DNA sequence populations. The method utilizes DNA-linked peptide substrates as activity probes. Signal detection involves chemical manipulation of a probe population downstream of sample exposure and application of purifying, selective pressure for enzyme products. Selection-induced changes in DNA abundance indicate sample activity. The detection of protein kinase, protease, and farnesyltransferase activities is demonstrated. The assays were employed to measure enzyme inhibition by small molecules and activity in cell lysates using parallel DNA sequencing or quantitative PCR. This strategy will allow the extensive infrastructure for genetic analysis to be applied to proteomic assays, which has a number of advantages in throughput, sensitivity, and sample multiplexing.

SUBMITTER: Jetson RR

PROVIDER: S-EPMC5402352 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature

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Sensing Enzymatic Activity by Exposure and Selection of DNA-Encoded Probes.

Jetson Rachael R RR Krusemark Casey J CJ

Angewandte Chemie (International ed. in English) 20160629 33

A sensing approach is applied to encode quantitative enzymatic activity information into DNA sequence populations. The method utilizes DNA-linked peptide substrates as activity probes. Signal detection involves chemical manipulation of a probe population downstream of sample exposure and application of purifying, selective pressure for enzyme products. Selection-induced changes in DNA abundance indicate sample activity. The detection of protein kinase, protease, and farnesyltransferase activitie ...[more]

PMID: 27355201

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