首页 | 本学科首页   官方微博 | 高级检索  
   检索      


Production of Functional Human Vitamin A Transporter/RBP Receptor (STRA6) for Structure Determination
Authors:Conor J Breen  Darren S Martin  Hui Ma  Kate McQuaid  Richard O’Kennedy  John B C Findlay
Institution:1. Department of Biology, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland.; 2. National Centre for Sensor Research, Biomedical Diagnostics Institute, Dublin City University, Dublin, Ireland.; Duke University, UNITED STATES,
Abstract:STRA6 is a plasma membrane protein that mediates the transport of vitamin A, or retinol, from plasma retinol binding protein (RBP) into the cell. Mutations in human STRA6 are associated with Matthew-Wood syndrome, which is characterized by severe developmental defects. Despite the obvious importance of this protein to human health, little is known about its structure and mechanism of action. To overcome the difficulties frequently encountered with the production of membrane proteins for structural determination, STRA6 has been expressed in Pichia pastoris as a fusion to green fluorescent protein (GFP), a strategy which has been a critical first step in solving the crystal structures of several membrane proteins. STRA6-GFP was correctly targeted to the cell surface where it bound RBP. Here we report the large-scale expression, purification and characterisation of STRA6-GFP. One litre of culture, corresponding to 175 g cells, yielded about 1.5 mg of pure protein. The interaction between purified STRA6 and its ligand RBP was studied by surface plasmon resonance-based binding analysis. The interaction between STRA6 and RBP was not retinol-dependent and the binding data were consistent with a transient interaction of 1 mole RBP/mole STRA6.
Keywords:
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号