干旱区生态环境知识资源中心(Arid): Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?

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DOI	10.1016/j.ibmb.2020.103392
	Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?
	Lismont, Els; Verbakel, Lina; Vogel, Elise; Corbisier, Jenny; Degroot, Gaetan-Nagim; Verdonck, Rik; Verlinden, Heleen; Marchal, Elisabeth; Springael, Jean-Yves; Vanden Broeck, Jozef
通讯作者	Verbakel, L
来源期刊	INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
ISSN	0965-1748
EISSN	1879-0240
出版年	2020
卷号	122
英文摘要	G protein-coupled receptors (GPCRs) are membrane-bound receptors that are considered prime candidates for the development of novel insect pest management strategies. However, the molecular signaling properties of insect GPCRs remain poorly understood. In fact, most studies on insect GPCR signaling are limited to analysis of fluctuations in the secondary messenger molecules calcium (Ca2+) and/or cyclic adenosine monophosphate (cAMP). In the current study, we characterized a corticotropin-releasing factor-related diuretic hormone (CRF-DH) receptor of the desert locust, Schistocerca gregaria. This Schgr-CRF-DHR is mainly expressed in the nervous system and in brain-associated endocrine organs. The neuropeptide Schgr-CRF-DH induced Ca2+-dependent aequorin-based bioluminescent responses in CHO cells co-expressing this receptor with the promiscuous G alpha(16) protein. Furthermore, when co-expressed with the cAMP-dependent bioluminescence resonance energy transfer (BRET)-based CAMYEL biosensor in HEK293T cells, this receptor elicited dose-dependent agonist-induced responses with an EC50 in the nanomolar range (4.02 nM). In addition, we tested if vertebrate BRET-based G protein biosensors, can also be used to detect direct G alpha protein subunit activation by an insect GPCR. Therefore, we analyzed ten different human BRET-based G protein biosensors, representing members of all four G alpha protein subfamilies; G alpha(s), G alpha(i/o), G alpha(q/11) and G alpha(12/13). Our data demonstrate that stimulation of Schgr-CRF-DHR by Schgr-CRF-DH can dose-dependently activate G alpha(i/o) and G alpha(s) biosensors, while no significant effects were observed with the G alpha(q/11) and G alpha(12/13) biosensors. Our study paves the way for future biosensor-based studies to analyze the signaling properties of insect GPCR5 in both fundamental science and applied research contexts.
英文关键词	BRET Cyclic AMP G protein-coupled receptor Insect Neuropeptide Reporter assay
类型	Article
语种	英语
开放获取类型	Green Published
收录类别	SCI-E
WOS记录号	WOS:000537570800012
WOS关键词	NEUROPEPTIDE-F RECEPTOR ; BLOOD-FEEDING BUG ; MALPIGHIAN TUBULES ; ALLATOTROPIN RECEPTOR ; COUPLED RECEPTORS ; MOLECULAR-CLONING ; GAMMA SUBUNITS ; HOUSE CRICKET ; TSETSE-FLY ; G-BETA
WOS类目	Biochemistry & Molecular Biology ; Entomology
WOS研究方向	Biochemistry & Molecular Biology ; Entomology
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/324622
作者单位	[Lismont, Els; Verbakel, Lina; Vogel, Elise; Verdonck, Rik; Verlinden, Heleen; Marchal, Elisabeth; Vanden Broeck, Jozef] Katholieke Univ Leuven, Mol Dev Physiol & Signal Transduct, Naamsestr 59,POB 02465, B-3000 Leuven, Belgium; [Corbisier, Jenny; Degroot, Gaetan-Nagim; Marchal, Elisabeth] IMEC, Kapeldreef 75, B-3001 Leuven, Belgium; [Springael, Jean-Yves] ULB, IRIBHM, Campus Erasme,808 Route Lennik, B-1070 Brussels, Belgium
推荐引用方式 GB/T 7714	Lismont, Els,Verbakel, Lina,Vogel, Elise,et al. Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?[J],2020,122.
APA	Lismont, Els.,Verbakel, Lina.,Vogel, Elise.,Corbisier, Jenny.,Degroot, Gaetan-Nagim.,...&Vanden Broeck, Jozef.(2020).Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?.INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY,122.
MLA	Lismont, Els,et al."Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?".INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY 122(2020).