Biotechnology Letters, Vol.28, No.19, 1581-1586, 2006
Expression and localization of recombinant human EDG-1 receptors in Pichia pastoris
The receptor for human endothelial differentiation gene-1 protein (EDG-1) was C-terminally tagged with green fluorescent protein and expressed in the methylotrophic yeast, Pichia pastoris. EDG-1 expression was driven by the highly inducible alcohol oxidase 1 promoter. Expression of EDG-1 recombinant protein was detected by Western blot analysis and confocal microscopy. The recombinant EDG-1 receptor protein was located in the plasma membrane. Radioligand binding assays demonstrated that the EDG-1 receptors expressed in Pichia pastoris have specific and saturation binding of P-32-labeled sphingosine 1-phosphate.
Keywords:endothelial differentiation gene;green fluorescent protein;heterologous expression;Pichia pastoris;sphingosine 1-phosphate