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Glycation of Glutamate Cysteine Ligase by 2-Deoxy-d-Ribose and its Potential Impact on Chemoresistance in Glioblastoma

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Abstract

The antioxidant glutathione (GSH) plays a critical role in maintaining intracellular redox homeostasis but in tumors the GSH biosynthetic pathway is often dysregulated, contributing to tumor resistance to radiation and chemotherapy. Glutamate-cysteine ligase (GCL) catalyzes the first and rate-limiting reaction in GSH synthesis, and enzyme function is controlled by GSH feedback inhibition or by transcriptional upregulation of the catalytic (GCLC) and modifier (GCLM) subunits. However, it has recently been reported that the activity of GCLC and the formation of GCL can be modified by reactive aldehyde products derived from lipid peroxidation. Due to the susceptibility of GCLC to posttranslational modifications by reactive aldehydes, we examined the potential for 2-deoxy-d-ribose (2dDR) to glycate GCLC and regulate enzyme activity and GCL formation. 2dDR was found to directly modify both GCLC and GCLM in vitro, resulting in a significant inhibition of GCLC and GCL enzyme activity without altering substrate affinity or feedback inhibition. 2dDR-mediated glycation also inhibited GCL subunit heterodimerization and formation of the GCL holoenzyme complex while not causing dissociation of pre-formed holoenzyme. This PTM could be of particular importance in glioblastoma (GBM) where intratumoral necrosis provides an abundance of thymidine, which can be metabolized by thymidine phosphorylase (TP) to form 2dDR. TP is expressed at high levels in human GBM tumors and shRNA knockdown of TP in U87 GBM cells results in a significant increase in cellular GCL enzymatic activity.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, 12850 East Montview Boulevard, V20-2102, Aurora, CO, 80045, USA

    Donald S. Backos, Kristofer S. Fritz, Debbie G. McArthur, Jadwiga K. Kepa, Dennis R. Petersen, Christopher C. Franklin & Philip Reigan

  2. Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    Andrew M. Donson & Nicholas K. Foreman

  3. University of Colorado Cancer Center, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    Nicholas K. Foreman, Christopher C. Franklin & Philip Reigan

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  1. Donald S. Backos
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  2. Kristofer S. Fritz
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  9. Philip Reigan
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Correspondence to Philip Reigan.

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Backos, D.S., Fritz, K.S., McArthur, D.G. et al. Glycation of Glutamate Cysteine Ligase by 2-Deoxy-d-Ribose and its Potential Impact on Chemoresistance in Glioblastoma. Neurochem Res 38, 1838–1849 (2013). https://doi.org/10.1007/s11064-013-1090-4

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