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Mol Genet Metab. 2015 Apr;114(4):570-9. doi: 10.1016/j.ymgme.2015.02.007. Epub 2015 Mar 5.

Consequences of impaired purine recycling on the proteome in a cellular model of Lesch-Nyhan disease.

Author information

  • 1Department of Biochemistry, Emory University, Atlanta, GA, USA.
  • 2Department of Neurology, Emory University, Atlanta, GA, USA.
  • 3Department of Biostatistics and Bioinformatics, Emory University, Atlanta, GA, USA.
  • 4Department of Neurology, Emory University, Atlanta, GA, USA; Department of Human Genetics & Pediatrics, Emory University, Atlanta, GA, USA. Electronic address: hjinnah@emory.edu.

Abstract

The importance of specific pathways of purine metabolism for normal brain function is highlighted by several inherited disorders, such as Lesch-Nyhan disease (LND). In this disorder, deficiency of the purine recycling enzyme, hypoxanthine-guanine phosphoribosyltransferase (HGprt), causes severe neurological and behavioral abnormalities. Despite many years of research, the mechanisms linking the defect in purine recycling to the neurobehavioral abnormalities remain unclear. In the current studies, an unbiased approach to the identification of potential mechanisms was undertaken by examining changes in protein expression in a model of HGprt deficiency based on the dopaminergic rat PC6-3 line, before and after differentiation with nerve growth factor (NGF). Protein expression profiles of 5 mutant sublines carrying different mutations affecting HGprt enzyme activity were compared to the HGprt-competent parent line using the method of stable isotopic labeling by amino acids in cell culture (SILAC) followed by denaturing gel electrophoresis with liquid chromatography and tandem mass spectrometry (LC-MS/MS) of tryptic digests, and subsequent identification of affected biochemical pathways using the Database for Annotation, Visualization and Integrated Discovery (DAVID) functional annotation chart analysis. The results demonstrate that HGprt deficiency causes broad changes in protein expression that depend on whether the cells are differentiated or not. Several of the pathways identified reflect predictable consequences of defective purine recycling. Other pathways were not anticipated, disclosing previously unknown connections with purine metabolism and novel insights into the pathogenesis of LND.

KEYWORDS:

Dopamine neurons; Hypoxanthine–guanine phosphoribosyltransferase; Lesch–Nyhan disease; Proteome

PMID:
25769394
PMCID:
PMC4390545
DOI:
10.1016/j.ymgme.2015.02.007
[PubMed - indexed for MEDLINE]
Free PMC Article
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