Expression, purification and phosphoinositide binding specifity of recombinant human SNX7 expressed in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1436-1445, 2014.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-345581
Responsible library:
WPRO
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.
Full text:
Available
Health context:
Neglected Diseases
Health problem:
Neglected Diseases
/
Zoonoses
Database:
WPRIM (Western Pacific)
Main subject:
Phosphatidylinositols
/
Substrate Specificity
/
Recombinant Proteins
/
Escherichia coli
/
Sorting Nexins
/
Genetic Vectors
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2014
Document type:
Article