VEGF internalization is not required for VEGFR-2 phosphorylation in bioengineered surfaces with covalently linked VEGF.
VEGF internalization is not required for VEGFR-2 phosphorylation in bioengineered surfaces with covalently linked VEGF.
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DOI：
10.1039/c1ib00037c
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发表时间：
2011-09
期刊：
影响因子：
--
通讯作者：
Segura T
中科院分区：
文献类型：
--
作者：
Anderson SM;Shergill B;Barry ZT;Manousiouthakis E;Chen TT;Botvinick E;Platt MO;Iruela-Arispe ML;Segura T
血管内皮生长因子(VEGF)通过其受体-2(VEGFR-2)的磷酸化激活内皮细胞的增殖、迁移和存活通路。血管内皮生长因子已通过包埋、静电隔离和共价结合等方法被引入生物材料中，但这些固定化策略对血管内皮生长因子信号转导途径的影响还没有得到深入的研究。此外，尽管生长因子与受体一起内化通常发生在生理环境中，但这种内化是否是受体磷酸化所必需的还不完全清楚。在这里，我们表明，通过修饰的肝素分子共价结合的血管内皮生长因子在人脐静脉内皮细胞(HUVECs)中引起pVEGFR-2的延长反应，并且在受体内吞过程中，共价连接减少了生长因子的内化。光钳测量表明，当VEGF和肝素之间引入共价键时，破坏肝素-血管内皮生长因子-VEGFR-2相互作用所需的破裂力从3-8pN增加到6-12pN。重要的是，通过将血管内皮生长因子与肝素底物共价结合，血管内皮生长因子的稳定性(半衰期)延长了三倍以上。在这里，数学模型支持生物学结论，进一步表明，当共价结合时，血管内皮生长因子的内化显著减少，并表明血管内皮生长因子可用于重复的磷酸化事件。
Vascular endothelial growth factor (VEGF) is known to activate proliferation, migration, and survival pathways in endothelial cells through phosphorylation of VEGF receptor-2 (VEGFR-2). VEGF has been incorporated into biomaterials through encapsulation, electrostatic sequestration, and covalent attachment, but the effect of these immobilization strategies on VEGF signaling has not been thoroughly investigated. Further, although growth factor internalization along with the receptor generally occurs in a physiological setting, whether this internalization is needed for receptor phosphorylation is not entirely clear. Here we show that VEGF covalently bound through a modified heparin molecule elicits an extended response of pVEGFR-2 in human umbilical vein endothelial cells (HUVECs) and that the covalent linkage reduces internalization of the growth factor during receptor endocytosis. Optical tweezer measurements show that the rupture force required to disrupt the heparin-VEGF-VEGFR-2 interaction increases from 3–8 pN to 6–12 pN when a covalent bond is introduced between VEGF and heparin. Importantly, by covalently binding VEGF to a heparin substrate, the stability (half-life) of VEGF is extended over three-fold. Here, mathematical models support the biological conclusions, further suggesting that VEGF internalization is significantly reduced when covalently bound, and indicating that VEGF is available for repeated phosphorylation events.
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DOI：
10.1083/jcb.200409115
发表时间：
2005-05-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lee S;Jilani SM;Nikolova GV;Carpizo D;Iruela-Arispe ML
通讯作者：
Iruela-Arispe ML
影响因子：
11.8
作者：
Lanahan, Anthony A.;Hermans, Karlien;Claes, Filip;Kerley-Hamilton, Joanna S.;Zhuang, Zhen W.;Giordano, Frank J.;Carmeliet, Peter;Simons, Michael
通讯作者：
Simons, Michael
DOI：
10.1073/pnas.072685299
发表时间：
2002-04-16
影响因子：
11.1
作者：
Hodneland, CD;Lee, YS;Mrksich, M
通讯作者：
Mrksich, M
影响因子：
4.8
作者：
Mellberg, Sofie;Dimberg, Anna;Claesson-Welsh, Lena
通讯作者：
Claesson-Welsh, Lena
影响因子：
14
作者：
Hosack, Luke W.;Firpo, Matthew A.;Peattie, Robert A.
通讯作者：
Peattie, Robert A.