Anti-angiogenic potential of trypsin inhibitor purified from Cucumis melo seeds: Homology modeling and molecular docking perspective

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• 作者：Hassan Rasouli^a ; Sharham Parvaneh^a ; Azadeh Mahnam^a ; Mohsen Rastegari-Pouyani^b ; Zohreh Hoseinkhani^a ; Kamran Mansouri^a ; ^{kmansouri@kums.ac.ir} ; ^{kamranmansouri@gmail.com}
• 关键词：BSA ; bovine serum albumin ; DMEM ; Dulbecco&rsquo ; s modified eagle&rsquo ; s medium ; BBI ; Bowman-Birk inhibitor ; ELISA ; enzyme-linked immunosorbent assay ; FBS ; fetal bovine serum ; HUVEC ; human umbilical vein endothelial cells ; LDH ; lactate dehydrogenase ; MTT ; 3-(4 ; 5-dimethylthiazol-2yl)-2 ; 5-diphenyl-2H-tetrazolium bromide ; PBS ; phosphate buffered saline ; PBST ; PBS containing tween 20 ; PDB ; protein data bank ; SDS-PAGE ; sodium dodecyl sulfate polyacrylamide gel electrophoresis ; TICMS ; trypsin inhibito
• 刊名：International Journal of Biological Macromolecules
• 出版年：2017
• 出版时间：March 2017
• 年：2017
• 卷：96
• 期：Complete
• 页码：118-128
• 全文大小：3694 K
• 卷排序：96

文摘

Melons have a good source of protease inhibitors. Its fruit and seeds have been used as a traditional medicine. However, its effects on angiogenesis and mechanism of its action remain elusive.Herein trypsin inhibitor from aqueous extract of C. melo seeds (TICMS) was purified. Its effects on different steps of angiogenesis were evaluated. Also, we examined its effects on migration and angiogenesis of endothelial cells. Three dimensional model of TICMS protein was accurately built in which TICMS docked to αVβ3 integrin and VEGFR1.Electrophoresis analysis of the purified protein revealed a single band with a molecular mass of about 3 kDa. Treatment with TICMS at six doses resulted in a significant decrease of endothelial cell proliferation with an IC50 value of about 20 μg/ml. Tubulogenesis assay revealed that a dose dependent anti-angiogenic activity of TICMS (5–40 μg/ml). Also, TICMS had inhibitory effects on VEGF, MMP-2 and MMP-9 secretion. Our docking result speculated that TICMS could bind to the cleft between the αVβ3 integrin and it able to decrease the activity of this receptor. The TICMS was also able to interact with VEGFR1 receptor, but with low probability. Based on our study, TICMS could be used as a specific angiogenesis inhibitor.