Revealing multi-binding sites for taspine to VEGFR-2 by cell membrane chromatography zonal elution.
A new high-expression vascular endothelial growth factor receptor-2 (VEGFR-2) cell membrane chromatography (CMC) method was developed to investigate the affinity of ligands for VEGFR-2. An HEK293 VEGFR-2/CMC system was applied to specifically recognize ligands acting on VEGFR-2. Sorafenib was used as a mobile phase additive to evaluate the effect of the marker's concentration on the retention of sorafenib and taspine, respectively. The relationship among the retention, the types of binding sites and the affinity of taspine binding to VEGFR-2 has also been concerned. The retention behavior indicated that sorafenib had two major binding regions on VEGFR-2, and that taspine might act as a multi-target VEGFR-2 inhibitor with similar biological activity to sorafenib. The equilibrium dissociation constants (K(D)) obtained from the model are (5.25 ± 0.31) × 10⁻⁷ and (9.88 ± 0.54) × 10⁻⁵ mol L⁻¹ for sorafenib at the high- and low-affinity sites, respectively, and the corresponding values for taspine are (3.88 ± 0.31) × 10⁻⁶ and (7.04 ± 0.49)×10⁻⁵ mol L⁻¹. The two types of binding sites contributed about a 1:2 ratio on the retention of taspine.
Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, China.
Hui
H
Du
Sicen
S
Wang
Jing
J
Ren
Nan
N
Lv
Langchong
L
He
D000470:Alkaloids / Q000737:chemistry* / Q000378:metabolism; D001557:Benzenesulfonates / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002845:Chromatography / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D009536:Niacinamide / Q000031:analogs & derivatives; D010671:Phenylurea Compounds; D011725:Pyridines / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D000077157:Sorafenib; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000737:chemistry* / Q000378:metabolism
D000470:Alkaloids; D001557:Benzenesulfonates; D010671:Phenylurea Compounds; D011725:Pyridines; D009536:Niacinamide; D000077157:Sorafenib; D040301:Vascular Endothelial Growth Factor Receptor-2; C018394:taspine