Work

Human phosphoglycerate dehydrogenase produces the oncometabolite D-2-hydroxyglutarate and promotes histone methylation

Year: 2014

Type: article

Abstract: Materials and methods To analyze the catalytic activity of PHGDH on possible alternative substrates, we conducted in vitro biochemical assays on recombinant human PHGDH. The reaction rate in the reductive direction with various substrates was monitored based on NADH consumption, measured by absorbance at 340 nm. The identity and chirality of products were analyzed by liquid chromatography-mass spectrometry (LC-MS) and by gas chromatography-mass spectrometry (GC-MS) after (R)-2-butanol derivatization. To analyze the intracellular function of PHGDH, we generated stable PHGDH knockdown cell lines, using two shRNA sequences. Metabolite levels in the knockdown cells were analyzed by LC-MS [4], and DNA and histone methylation was analyzed by immunoblotting.

Source: Cancer & Metabolism

DOI: 10.1186/2049-3002-2-s1-p75

Authors: Jing Fan, Xin Teng, Ling Liu, Ryan Looper, Joshua Rabinowitz

Institutions: Princeton University, University of Utah, Rutgers, The State University of New Jersey, Rutgers Cancer Institute

Language: English

Topic: Cancer, Hypoxia, and Metabolism

Subfield: Cancer Research

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $3,190