One-Pot Chemoenzymatic Cascade for Labeling of the Epigenetic Marker 5-Hydroxymethylcytosine

Gil Nifker, Michal Levy-Sakin, Yifat Berkov-Zrihen, Tamar Shahal, Tslil Gabrieli, Micha Fridman, Yuval Ebenstein

Research output: Contribution to journalArticlepeer-review


The epigenetic DNA modification 5-hydroxymethylcytosine (5-hmC) is important for the regulation of gene expression during development and in tumorigenesis. 5-hmC can be selectively glycosylated by T4 β-glucosyltransferase (β-GT); introduction of an azide on the attached sugar provides a chemical handle for isolation or fluorescent tagging of 5-hmC residues by click chemistry. This approach has not been broadly adopted because of the challenging synthesis and limited commercial availability of the glycosylation substrate, 6-deoxy-6-azido-α-D-glucopyranoside. We report the enzyme-assisted synthesis of this precursor by the uridylyltransferase from Pasteurella multocida (PmGlmU). We were able to directly label 5-hmC in genomic DNA by an enzymatic cascade involving successive action of PmGlmU and β-GT. This is a facile and cost-effective one-pot chemoenzymatic methodology for 5-hmC analysis.

Original languageEnglish
Pages (from-to)1857-1860
Number of pages4
Issue number13
StatePublished - 1 Sep 2015


  • DNA methylation
  • chemoenzymatic synthesis
  • fluorescent probes
  • hydroxymethylcytosine
  • single molecules
  • sugar nucleotides


Dive into the research topics of 'One-Pot Chemoenzymatic Cascade for Labeling of the Epigenetic Marker 5-Hydroxymethylcytosine'. Together they form a unique fingerprint.

Cite this