Identification of proteins interacting with GTP cyclohydrolase I

作者：Du Jianhai; Xu Hao; Wei Na; Wakim Bassam; Halligan Brian; Pritchard Kirkwood A Jr; Shi Yang^*

来源：Biochemical and Biophysical Research Communications, 2009, 385(2): 143-147.

DOI：10.1016/j.bbrc.2009.05.026

• GTP cyclohydrolase I
• Proteomic
• Interacting proteins
• Tetrahydrobiopterin
• Nitric oxide synthase
• Aromatic amino acid hydroxylase

GTP cyclohydrolase I (GCH-1) is the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin, all 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.

• 出版日期2009-7-24
• 单位山东大学; 中国医科大学