Absorptive-Mediated Endocytosis of an Adrenocorticotropic Hormone (ACTH) Analogue, Ebiratide, into the Blood–Brain Barrier: Studies with Monolayers of Primary Cultured Bovine Brain Capillary Endothelial Cells
Absorptive-Mediated Endocytosis of an Adrenocorticotropic Hormone (ACTH) Analogue, Ebiratide, into the Blood–Brain Barrier: Studies with Monolayers of Primary Cultured Bovine Brain Capillary Endothelial Cells
复制标题
促肾上腺皮质激素 (ACTH) 类似物依比拉肽吸收介导的内吞作用进入血脑屏障：原代培养牛脑毛细血管内皮细胞单层的研究
作者：
T. Terasaki;S. Takakuwa;A. Saheki;S. Moritani;T. Shimura;S. Tabata;A. Tsuji
本文研究了神经调节性促肾上腺皮质激素类似物[125I](H-Met(O2)-Glu[125I]His-Phe-D-Lys-Phe-NH(CH2)8NH2)，在培养的牛脑毛细血管内皮细胞中的内化作用。孵育液的高效液相色谱分析表明，在与BCEC孵育的过程中，[125I]Euratide没有代谢。[~(125)I]Euratide与BCEC的耐酸结合力随时间的延长而增加，且与温度和介质渗透压有明显的依赖关系。丹西卡维林或氧化苯砷、内吞抑制剂和代谢抑制剂2，4-二硝基苯酚可显著降低[125I]EBiratide的耐酸结合率。在未标记的伊比拉肽(100 nM-1 mm)存在下，[125I]伊比拉肽的耐酸结合是饱和的。30min时最大内化容量(Bmax)为7.96±3.27微克分子/毫克蛋白质，半饱和常数(Kd)为15.9±6.4微米。碱性多肽如多L赖氨酸、鱼精蛋白、组蛋白和促肾上腺皮质激素可抑制耐酸结合，而多L谷氨酸、胰岛素和转铁蛋白不能抑制其耐酸结合。这些结果证实，伊比拉肽是通过吸收介导的内吞作用通过血脑屏障转运的。
The internalization of a neuromodulatory adrenocorticotropic hormone (ACTH) analogue, [125I]ebiratide (H-Met(O2)-Glu[125I]His-Phe-D-Lys-Phe-NH(CH2)8NH2), was examined in cultured mono-layers of bovine brain capillary endothelial cells (BCEC). HPLC analysis of the incubation solution showed that [125I]ebiratide was not metabolized during the incubation with BCEC. The acid-resistant binding of [125I]ebiratide to BCEC increased with time for 120 min and showed a significant dependence on temperature and medium osmolarity. Pretreatment of BCEC with dansylcadaverine or phenylarsine oxide, endocytosis inhibitors, and 2,4-dinitrophenol, a metabolic inhibitor, decreased significantly the acid-resistant binding of [125I]ebiratide. The acid-resistant binding of [125I]ebiratide was saturable in the presence of unlabeled ebiratide (100 nM–1 mM). The maximal internalization capacity (Bmax) at 30 min was 7.96 ± 3.27 µmol/mg of protein with a half-saturation constant (Kd) of 15.9 ± 6.4 µM. The acid-resistant binding was inhibited by basic peptides such as poly-L-lysine, protamine, histone, and ACTH but was not inhibited by poly-L-glutamic acid, insulin, or transferrin. These results confirmed that ebiratide is transported through the blood-brain barrier via an absorptive-mediated endocytosis.
影响因子：
9.8
作者：
PARDRIDGE, WM;EISENBERG, J;JING, Y
通讯作者：
JING, Y
细胞表面胰岛素受体快速、可逆的内化。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Knutson,VP;Ronnett,GV;Lane,MD
通讯作者：
Lane,MD