کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10814768 | 1058397 | 2016 | 46 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
FRET biosensors reveal AKAP-mediated shaping of subcellular PKA activity and a novel mode of Ca2Â +/PKA crosstalk
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کلمات کلیدی
IBMXCB4ECFPAKAPβ2AREYFPforskolinGPCRFsknESeGFPCFP3-isobutyl-1-methylxanthine - 3-ایزوبوتیل-1-متیلکسانتینcAMP - cAMPG protein coupled receptor - G پروتئین همراه با پروتئین[Ca2 +]i - [Ca2 +] iCyclic adenosine monophosphate - آدنوزین مونوفسفات Cyclicenhanced cyan fluorescent protein - افزایش پروتئین فلورسنت سیانوژنFRET - انتقال انرژی رزونانسی فورسترISO - ایزوisoproterenol - ایزوپروترنولCAM - ساخت به کمک کامپیوترnuclear export signal - سیگنال صادرات هسته ایintracellular calcium concentration - غلظت کلسیم داخل سلولیA-kinase anchoring protein - پروتئین anchoring A-kinaseenhanced yellow fluorescent protein - پروتئین فلورسنت زرد افزایش یافته استenhanced green fluorescent protein - پروتئین فلورسنت سبز افزایش یافته استcyan fluorescent protein - پروتئین فلورسنت سیانوژنcalcium-calmodulin - کلسیم کلسیم کلسیمkilodaltons - کیلودالتونβ2-adrenergic receptor - گیرنده β2-adrenergic
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Scaffold proteins play a critical role in cellular homeostasis by anchoring signaling enzymes in close proximity to downstream effectors. In addition to anchoring static enzyme complexes, some scaffold proteins also form dynamic signalosomes that can traffic to different subcellular compartments upon stimulation. Gravin (AKAP12), a multivalent scaffold, anchors PKA and other enzymes to the plasma membrane under basal conditions, but upon [Ca2 +]i elevation, is rapidly redistributed to the cytosol. Because gravin redistribution also impacts PKA localization, we postulate that gravin acts as a calcium “switch” that modulates PKA-substrate interactions at the plasma membrane, thus facilitating a novel crosstalk mechanism between Ca2 + and PKA-dependent pathways. To assess this, we measured the impact of gravin-V5/His expression on compartmentalized PKA activity using the FRET biosensor AKAR3 in cultured cells. Upon treatment with forskolin or isoproterenol, cells expressing gravin-V5/His showed elevated levels of plasma membrane PKA activity, but cytosolic PKA activity levels were reduced compared with control cells lacking gravin. This effect required both gravin interaction with PKA and localization at the plasma membrane. Pretreatment with calcium-elevating agents thapsigargin or ATP caused gravin redistribution away from the plasma membrane and prevented gravin from elevating PKA activity levels at the membrane. Importantly, this mode of Ca2 +/PKA crosstalk was not observed in cells expressing a gravin mutant that resisted calcium-mediated redistribution from the cell periphery. These results reveal that gravin impacts subcellular PKA activity levels through the spatial targeting of PKA, and that calcium elevation modulates downstream β-adrenergic/PKA signaling through gravin redistribution, thus supporting the hypothesis that gravin mediates crosstalk between Ca2 + and PKA-dependent signaling pathways. Based on these results, AKAP localization dynamics may represent an important paradigm for the regulation of cellular signaling networks.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Cellular Signalling - Volume 28, Issue 4, April 2016, Pages 294-306
Journal: Cellular Signalling - Volume 28, Issue 4, April 2016, Pages 294-306
نویسندگان
Micah B. Schott, Faith Gonowolo, Benjamin Maliske, Bryon Grove,