Details

Autor(en) / Beteiligte

• Pals, Mathijs J.
• Velema, Willem A.

Titel

A Quenched Size‐Expanded Nucleotide Reports Activity of the Leukemia Biomarker Terminal Deoxynucleotidyl Transferase (TdT)

Ist Teil von

• Angewandte Chemie International Edition, 2023-04, Vol.62 (18), p.e202302796-n/a

Auflage

International ed. in English

Ort / Verlag

Germany: Wiley Subscription Services, Inc

Erscheinungsjahr

2023

Quelle

MEDLINE

Beschreibungen/Notizen

• Terminal deoxynucleotidyl Transferase (TdT) is a template‐independent DNA polymerase that plays an essential role in the human adaptive immune system and is upregulated in several types of leukemia. It has therefore gained interest as a leukemia biomarker and potential therapeutic target. Herein, we describe a FRET‐quenched fluorogenic probe based on a size‐expanded deoxyadenosine that reports directly on TdT enzymatic activity. The probe enables real‐time detection of primer extension and de novo synthesis activity of TdT and displays selectivity over other polymerase and phosphatase enzymes. Importantly, TdT activity and its response to treatment with a promiscuous polymerase inhibitor could be monitored in human T‐lymphocyte cell extract and Jurkat cells using a simple fluorescence assay. Finally, employing the probe in a high‐throughput assay resulted in the identification of a non‐nucleoside TdT inhibitor. A fluorogenic probe based on size‐expanded adenosine is used to monitor activity of the leukemia biomarker and therapeutic target Terminal deoxynucleotidyl Transferase (TdT). The probe can measure TdT activity in cancer cell extract in real‐time and report on its response to treatment. Use of the probe in a high‐throughput screening led to the identification of the non‐nucleoside inhibitor SCH202676.