Identification of proteins interacting with GTP cyclohydrolase I. Biochem Biophys Res Commun 2009 Jul 24;385(2):143-7
Date
05/16/2009Pubmed ID
19442649Pubmed Central ID
PMC2737525DOI
10.1016/j.bbrc.2009.05.026Scopus ID
2-s2.0-67349116808 (requires institutional sign-in at Scopus site) 9 CitationsAbstract
GTP cyclohydrolase I (GCH-1) is the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin, an essential cofactor for nitric oxide synthase and aromatic amino acid hydroxylase. To explore the interactome of GCH-1, we established a HEK 293 cell line stably expressing tetracycline-inducible FLAG-GCH-1. FLAG-GCH-1 and associated proteins were immunoprecipitated and analyzed by liquid chromatography/tandem mass spectrometry. Twenty-nine proteins, derived from different subcellular components such as cytosol, membranes, nucleus and mitochondria were identified to interact with GCH-1. Cell fractionation studies also showed that GCH-1 was present in the cytosol, membranes and nucleus. Gene ontology analysis revealed that GCH-1 interactome was involved in a variety of biological processes such as signal transduction, apoptosis, metabolism, transport and cell organization. To our knowledge, this study is the first to provide a comprehensive analysis of the GCH-1 interactome. Findings expand the number and diversity of proteins that are known to associate with GCH-1.
Author List
Du J, Xu H, Wei N, Wakim B, Halligan B, Pritchard KA Jr, Shi YAuthor
Kirkwood A. Pritchard PhD Professor in the Surgery department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Cell FractionationCell Line
Cell Nucleus
Chromatography, Liquid
Cytosol
GTP Cyclohydrolase
Humans
Mitochondria
Proteins
Tandem Mass Spectrometry
