Display options
Cite
Breski M, Dey D, Obringer S, et al. Engineering Biological C-H Functionalization Leads to Allele-Specific Regulation of Histone Demethylases. J Am Chem Soc. 2016;138(41):13505-13508doi: 10.1021/jacs.6b08653.
Breski, M., Dey, D., Obringer, S., Sudhamalla, B., & Islam, K. (2016). Engineering Biological C-H Functionalization Leads to Allele-Specific Regulation of Histone Demethylases. Journal of the American Chemical Society, 138(41), 13505-13508. https://doi.org/10.1021/jacs.6b08653
Breski, Megan, et al. "Engineering Biological C-H Functionalization Leads to Allele-Specific Regulation of Histone Demethylases." Journal of the American Chemical Society vol. 138,41 (2016): 13505-13508. doi: https://doi.org/10.1021/jacs.6b08653
Breski M, Dey D, Obringer S, Sudhamalla B, Islam K. Engineering Biological C-H Functionalization Leads to Allele-Specific Regulation of Histone Demethylases. J Am Chem Soc. 2016 Oct 19;138(41):13505-13508. doi: 10.1021/jacs.6b08653. Epub 2016 Oct 10. PMID: 27709909.
Copy Download .nbib Format: NLM
Share it on

J Am Chem Soc. 2016 Oct 19;138(41):13505-13508. doi: 10.1021/jacs.6b08653. Epub 2016 Oct 10.

Engineering Biological C-H Functionalization Leads to Allele-Specific Regulation of Histone Demethylases.

Journal of the American Chemical Society

Megan Breski, Debasis Dey, Sara Obringer, Babu Sudhamalla, Kabirul Islam

Affiliations

  1. Department of Chemistry and ?Department of Human Genetics, University of Pittsburgh , Pittsburgh, Pennsylvania 15260, United States.

PMID: 27709909 DOI: 10.1021/jacs.6b08653

Abstract

Oxidative C-H hydroxylation of methyl groups, followed by their removal from DNA, RNA, or histones, is an epigenetic process critical to transcriptional reprogramming and cell fate determination. This reaction is catalyzed by Fe(II)-dependent dioxygenases using the essential metabolite 2-ketoglutarate (2KG) as a cofactor. Given that the human genome encodes for more than 60 2KG-dependent dioxygenases, assigning their individual functions remains a significant challenge. Here we describe a protein-ligand interface engineering approach to break the biochemical degeneracy of these enzymes. Using histone lysine demethylase 4 as a proof-of-concept, we show that the enzyme active site can be expanded to employ bulky 2KG analogues that do not sensitize wild-type demethylases. We establish the orthogonality, substrate specificity, and catalytic competency of the engineered demethylation apparatus in biochemical assays. We further demonstrate demethylation of cognate substrates in physiologically relevant settings. Our results provide a paradigm for rapid and conditional manipulation of histone demethylases to uncloak their isoform-specific functions.

Publication Types