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Mapping Post-Translational Modifications of de Novo Purine Biosynthetic Enzymes: Implications for Pathway Regulation
Journal article   Peer reviewed

Mapping Post-Translational Modifications of de Novo Purine Biosynthetic Enzymes: Implications for Pathway Regulation

Chunliang Liu, Giselle M. Knudsen, Anthony M. Pedley, Jingxuan He, Jared L. Johnson, Tomer M. Yaron, Lewis C. Cantley and Stephen J. Benkovic
Journal of proteome research, Vol.18(5), pp.2078-2087
05/03/2019
DOI: 10.1021/acs.jproteome.8b00969
PMCID: PMC6499638
PMID: 30964683
url
https://www.ncbi.nlm.nih.gov/pmc/articles/6499638View
Open Access

Abstract

Purines represent a class of essential metabolites produced by the cell to maintain cellular homeostasis and facilitate cell proliferation. In times of high purine demand, the de novo purine biosynthetic pathway is activated; however, the mechanisms that facilitate this process are largely unknown. One plausible mechanism is through intracellular signaling, which results in enzymes within the pathway becoming post-translationally modified to enhance their individual enzyme activities and the overall pathway metabolic flux. Here, we employ a proteomic strategy to investigate the extent to which de novo purine biosynthetic pathway enzymes are post-translationally modified in 293T cells. We identified 7 post-translational modifications on 135 residues across the 6 human pathway enzymes. We further asked whether there were differences in the post-translational modification state of each pathway enzyme isolated from cells cultured in the presence or absence of purines. Of the 174 assigned modifications, 67% of them were only detected in one experimental growth condition in which a significant number of serine and threonine phosphorylations were noted. A survey of the most-probable kinases responsible for these phosphorylation events uncovered a likely AKT phosphorylation site at residue Thr397 of PPAT, which was only detected in cells under purine-supplemented growth conditions. These data suggest that this modification might alter enzyme activity or modulate its interaction(s) with downstream pathway enzymes. Together, these findings propose a role for post-translational modifications in pathway regulation and activation to meet intracellular purine demand.
Biochemical Research Methods Biochemistry & Molecular Biology Life Sciences & Biomedicine Science & Technology

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