An Improved Targeted cAMP Sensor to Study the Regulation of Adenylyl Cyclase 8 by Ca2+ Entry through Voltage-Gated Channels

被引：27

作者：

Everett, Katy L. ^{[1
]}

Cooper, Dermot M. F. ^{[1
]}

机构：

[1] Univ Cambridge, Dept Pharmacol, Cambridge CB2 1QJ, England

来源：

PLOS ONE | 2013年 / 8卷 / 09期

基金：

英国惠康基金;

关键词：

PROTEIN-KINASE-A; PULSATILE INSULIN-SECRETION; YELLOW FLUORESCENT PROTEIN; SINGLE BETA-CELLS; DIFFERENT ISOFORMS; SIGNALING COMPLEX; CALCIUM-CHANNELS; LIPID RAFTS; CYCLIC-AMP; GLUCOSE;

D O I：

10.1371/journal.pone.0075942

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Here we describe an improved sensor with reduced pH sensitivity tethered to adenylyl cyclase (AC) 8. The sensor was used to study cAMP dynamics in the AC8 microdomain of MIN6 cells, a pancreatic beta-cell line. In these cells, AC8 was activated by Ca2+ entry through L-type voltage-gated channels following depolarisation. This activation could be reconstituted in HEK293 cells co-expressing AC8 and either the alpha 1C or alpha 1D subunit of L-type voltage-gated Ca2+ channels. The development of this improved sensor opens the door to the study of cAMP microdomains in excitable cells that have previously been challenging due to the sensitivity of fluorescent proteins to pH changes.

引用

页数：11

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