title: [Expression, purification and phosphoinositide binding specifity of recombinant human SNX7 expressed in Escherichia coli].
abstract: Sorting nexins (SNXs) are a large group of proteins that contain Phox (PX) domain and involve in regulating endocytosis and endosome sorting. SNX7, a member of SNXs family, contains a PX domain and a BAR domain. In zebrafish, SNX7 is a liver-enriched anti-apoptotic protein and indispensible for the liver development. A fragment of SNX7 cDNA ((px-bar)snx7), encoding the PX domain and the BAR domain, was inserted into the expressing vector p28a, transformed into E. coli Rosseta 2 (DE3), and then induced by isopropyl β-D-1-Thiogalactopyranoside (IPTG). After affinity, ion exchange and gel filtration purification, the purity of (PX-BAR)SNX7 reached over 95%. Dynamic light scattering (DLS) experiment indicated that (PX-BAR)SNX7 was homogeneous in solution. Lipid overlay assay showed that (PX-BAR)SNX7 can bind to PtdIns(5)P, PtdIns(4,5)P2 and PtdIns(3,4,5)P3.
authors.forename: Zhan
authors.initials: Z
authors.lastname: Feng
authors.forename: Tingting
authors.initials: T
authors.lastname: Xu
authors.forename: Jinxin
authors.initials: J
authors.lastname: Xu
mesh_terms: D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D010716:Phosphatidylinositols / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D058305:Sorting Nexins / Q000096:biosynthesis*; D013379:Substrate Specificity
chemical_list: D010716:Phosphatidylinositols; D011994:Recombinant Proteins; D058305:Sorting Nexins