Regulation of monocarboxylic acid transporter-1 by cAMP dependent vesicular trafficking in brain microvascular endothelial cells.
Regulation of monocarboxylic acid transporter-1 by cAMP dependent vesicular trafficking in brain microvascular endothelial cells.
复制标题
DOI：
10.1371/journal.pone.0085957
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Smith JP
中科院分区：
文献类型：
--
作者：
Uhernik AL;Li L;LaVoy N;Velasquez MJ;Smith JP
在这项研究中，对培养的大鼠脑微血管内皮细胞细胞质囊泡中的单羧酸转运体-1 （Mct1）进行了详细的表征，表明它们是一种多样化的核内体，通过短暂暴露于膜渗透的cAMP类似物8Br-cAMP 25至30分钟来调节转运体。囊泡的大小、流动性、内部pH值均不均匀，并与特定类型的核内体共定位，包括早期核内体、网格蛋白包被的囊泡、腔泡、反式高尔基体和溶酶体。Mct1的囊泡定位不依赖于其N或C末端，然而，Mct1囊泡的大小和pH值通过删除任何一端而增加，这表明该末端在Mct1的囊泡运输中起作用。使用本研究开发的一种基于BCECF-AM的新型检测方法，8Br-cAMP在25分钟后降低Mct1囊泡的pH值。这一结果和方法在比例ph敏感EGFP-mCherry双标记Mct1构建物的实验中得到了证实。总体而言，研究结果表明，cAMP信号通路通过促进Mct1进入一个高度动态的囊泡运输途径，从而降低了脑血管内皮细胞中Mct1的功能，该途径似乎导致转运蛋白运输到自噬体和溶酶体。
In this study, a detailed characterization of Monocarboxylic Acid Transporter-1 (Mct1) in cytoplasmic vesicles of cultured rat brain microvascular endothelial cells shows them to be a diverse population of endosomes intrinsic to the regulation of the transporter by a brief 25 to 30 minute exposure to the membrane permeant cAMP analog, 8Br-cAMP. The vesicles are heterogeneous in size, mobility, internal pH, and co-localize with discreet markers of particular types of endosomes including early endosomes, clathrin coated vesicles, caveolar vesicles, trans-golgi, and lysosomes. The vesicular localization of Mct1 was not dependent on its N or C termini, however, the size and pH of Mct1 vesicles was increased by deletion of either terminus demonstrating a role for the termini in vesicular trafficking of Mct1. Using a novel BCECF-AM based assay developed in this study, 8Br-cAMP was shown to decrease the pH of Mct1 vesicles after 25 minutes. This result and method were confirmed in experiments with a ratiometric pH-sensitive EGFP-mCherry dual tagged Mct1 construct. Overall, the results indicate that cAMP signaling reduces the functionality of Mct1 in cerebrovascular endothelial cells by facilitating its entry into a highly dynamic vesicular trafficking pathway that appears to lead to the transporter's trafficking to autophagosomes and lysosomes.
登录
查看更多内容
影响因子：
64.5
作者：
Huttlin EL;Jedrychowski MP;Elias JE;Goswami T;Rad R;Beausoleil SA;Villén J;Haas W;Sowa ME;Gygi SP
通讯作者：
Gygi SP
DOI：
10.1111/j.1600-0854.2010.01155.x
发表时间：
2011-04
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Castorino JJ;Deborde S;Deora A;Schreiner R;Gallagher-Colombo SM;Rodriguez-Boulan E;Philp NJ
通讯作者：
Philp NJ
影响因子：
5.6
作者：
ROUX, F;DURIEUTRAUTMANN, O;COURAUD, PO
通讯作者：
COURAUD, PO
DOI：
10.1161/01.atv.0000070546.16946.3a
发表时间：
2003-07-01
影响因子：
8.7
作者：
Frank, PG;Woodman, SE;Lisanti, MP
通讯作者：
Lisanti, MP
影响因子：
4.8
作者：
Smith, JP;Drewes, LR
通讯作者：
Drewes, LR