title: Expression and localization of recombinant human EDG-1 receptors in Pichia pastoris.
abstract: 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 (32)P-labeled sphingosine 1-phosphate.
authors.affiliation: Research Centre of Gene Diversity and Designed Agriculture, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Science, Fudan University, Shanghai 200433, P.R. China.
authors.forename: Guangxiao
authors.initials: G
authors.lastname: Yang
authors.forename: Tianlei
authors.initials: T
authors.lastname: Liu
authors.forename: Wulin
authors.initials: W
authors.lastname: Peng
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Sun
authors.forename: Han
authors.initials: H
authors.lastname: Zhang
authors.forename: Chaoqun
authors.initials: C
authors.lastname: Wu
authors.forename: Daleng
authors.initials: D
authors.lastname: Shen
mesh_terms: D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D010843:Pichia / Q000235:genetics*; D011869:Radioligand Assay; D049349:Receptors, Lysosphingolipid / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D049349:Receptors, Lysosphingolipid; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins