[Effect of N-terminal deletion on biological activity of vascular endothelial cell growth inhibitor].
Vascular endothelial cell growth inhibitor (VEGI) is a novel cytokine which belongs to the TNF superfamily. It can inhibit the proliferation of endothelial cells and neovascularization. However, little is known about the structure-function relationship of VEGI. In order to study the effect of the N-terminus of VEGI on biological properties, the sequence alignment among VEGI and TNF superfamily members based on structure knowledge was done, and then two truncated forms of VEGI were constructed, in which 43 and 51 amino acids from N-terminus were deleted and named VEGI(131) and VEGI(123). Recombinant proteins were generated from E. coli. The expression rates were 25.2% (VEGI(131)) and 27.8% (VEGI(123)) of total bacterial proteins. After purification the purity reached 92.5% (VEGI(131)) and 91.6% (VEGI(123)). VEGI(131) showed significant inhibitory effect on growth of human umbilical vein endothelial cells (HUVEC), IC(50) of VEGI(131) being 35 mg/L. Under the same conditions, IC(50) of VEGI(151) (the wild type of VEGI) was 27 mg/L, but VEGI(123) showed no inhibitory effect. On chick choriallantic membrane (CAM) assay, VEGI(151) markedly reduced the number of main vessels, and VEGI131 decreased capillary number, while the effect of VEGI(123) was almost the same as control. These results suggest that the first 43 amino acids from N-terminus of VEGI have no significant effect on biological activity, but the amino acids 44-51 at N-terminus are required for full biological activity.
Zhang
Min
M
Department of Microbiology, Second Military Medical University, Shanghai 200433, China.
Wang
Lu
L
Wang
Hong-Wei
HW
Pan
Xin
X
Pan
Wei
W
Qi
Zhong-Tian
ZT
D000482:Allantois / Q000098:blood supply; D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002642:Chick Embryo; D002823:Chorion / Q000098:blood supply; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D018919:Neovascularization, Physiologic / Q000187:drug effects; D016415:Sequence Alignment; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000494:pharmacology*
C117026:TNFSF15 protein, human; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15; D014409:Tumor Necrosis Factor-alpha