Author  
Place of duty  
Title   혈관신생 억제를 위한 가용성 변이형 VEGF 수용체의 제조 ( Production of Soluble VEGF Receptor Mutants for Inhibition of Angiogenesis )
Publicationinfo   2000 Jan; 032(03): 595-605.
Key_word   Angiogenesis , Vascular endothelial growth factor , Baculovirus , FLT - 1 , Tyrosine kinase , Mutant receptor
Full-Text  
Abstract   Purpose: Vascular endothelial growth factor (VEGF) is a potent angiogenic factor of many solid tumors, promoting vascularization and formation of metastases. In an attempt to generate effective VEGF inhibitors, the authors constructed the VEGF receptor mutants, expressed in E. coli and Sf9 insect cells, and examined their binding to VEGF. Materials and Methods: The cDNA fragment encoding FLT-1 extracellular domain was cloned from human umbilical vein endothelial (HUVE) cell total RNA using RT-PCR. PCR-subcloning was performed using this template, in order to generate the deletion mutants by introducing FLT-1 partial sequences into E.coli expression vector pET-21d and baculovirus transfer vactors, pBAC-1 and pBAC-3. Two mutant proteins from baculovirus-infected insect cells were purified by heparin sepharose chromatography and immobilized into nitrocellulose membrane followed by 125 I-VEGF binding assay. Results: Two mutant receptors, sFLT (1~7) and sFLT (2~4) expressed in E.coli appeared in inclusion body as insoluble proteins. The soluble mutant receptors were produced in low yield by baculovirus/insect cell expression system. Both immobilized mutant receptors, sFLT (1~7) and sFLT (2~4) were able to bind VEGF. Conclusion: These results suggest that a small soluble mutant receptor, sFLT (2~4), as well as sFLT (1~7) may be used effectively for blocking angiogenic function of VEGF.
저 자   윤수영(Soo Young Yun),홍용길(Yong Kil Hong),이윤(Yoon Lee),김광세(Kwang Sei Kim),김훈교(Hoon Kyo Kim),조영애(Young Ae Joe)