Work

Catalytic Space Engineering as a Strategy to Activate C–H Oxidation on 5-Methylcytosine in Mammalian Genome

Year: 2021

Type: article

Abstract: Conditional remodeling of enzyme catalysis is a formidable challenge in protein engineering. Herein, we have undertaken a unique active site engineering tactic to command catalytic outcomes. With ten-eleven translocation (TET) enzyme as a paradigm, we show that variants with an expanded active site significantly enhance multistep C-H oxidation in 5-methylcytosine (5mC), whereas a crowded cavity leads to a single-step catalytic apparatus. We further identify an evolutionarily conserved residue in the TET family with a remarkable catalysis-directing ability. The activating variant demonstrated its prowess to oxidize 5mC in chromosomal DNA for potentiating expression of genes including tumor suppressors.

Source: Journal of the American Chemical Society

DOI: 10.1021/jacs.1c03815

Authors: Sushma Sappa, Debasis Dey, Babu Sudhamalla, Kabirul Islam

Institution: University of Pittsburgh

Language: English

Topic: Metal-Catalyzed Oxygenation Mechanisms

Subfield: Inorganic Chemistry

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: National Science Foundation, National Institute of General Medical Sciences

Awards: Crosslinking-Assisted Substrate Identification for Lysine Demethylases, Protein-ligand interface engineering for allele-specific regulation of histone demethylases and epigenome editing, Site-Specific Photochemistry on Epigenetic Readers for Interactome Profiling