A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function

doi:10.3389/fphys.2019.00392

	A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function
	Ziraldo, Gaia 1,2; Buratto, Damiano3 ; Kuang, Yuanyuan3 ; Xu, Liang3,6 ; Carrer, Andrea 1,4; Nardin, Chiara 1,4; Chiani, Francesco 1; Salvatore, Anna Maria 1; Paludetti, Gaetano 2,5; Lerner, Richard A.3 ; Yang, Guang3 ; Zonta, Francesco3 ; Mammano, Fabio1,3,4
	2019-06-11
发表期刊	FRONTIERS IN PHYSIOLOGY
ISSN	1664-042X
卷号	10
发表状态	已发表
DOI	10.3389/fphys.2019.00392
摘要	Connexin hemichannels, which are plasma membrane hexameric channels (connexons) composed of connexin protein protomers, have been implicated in a host of physiological processes and pathological conditions. A number of single point pathological mutations impart a "leaky" character to the affected hemichannels, i. e., make them more active or hyperactive, suggesting that normal physiological condition could be recovered using selective hemichannel inhibitors. Recently, a human-derived monoclonal antibody named abEC1.1 has been shown to inhibit both wild type and hyperactive hemichannels composed of human (h) connexin 26 (hCx26) subunits. The aims of this work were (1) to characterize further the ability of abEC1.1 to selectively modulate connexin hemichannel function and (2) to assess its in vitro stability in view of future translational applications. In silico analysis of abEC1.1 interaction with the hCx26 hemichannel identified critically important extracellular domain amino acids that are conserved in connexin 30 (hCx30) and connexin 32 (hCx32). Patch clamp experiments performed in HeLa DH cells confirmed the inhibition efficiency of abEC1.1 was comparable for hCx26, hCx30 and hCx32 hemichannels. Of note, even a single amino acid difference in the putative binding region reduced drastically the inhibitory effects of the antibody on all the other tested hemichannels, namely hCx30.2/31.3, hCx30.3, hCx31, hCx31.1, hCx37, hCx43 and hCx45. Plasma membrane channels composed of pannexin 1 were not affected by abEC1.1. Finally, size exclusion chromatography assays showed the antibody does not aggregate appreciably in vitro. Altogether, these results indicate abEC1.1 is a promising tool for further translational studies.
关键词	connexin hemichannels rare diseases phage display libraries therapeutic monoclonal antibodies molecular dynamics patch clamp
收录类别	SCI ; SCIE
语种	英语
资助项目	Science and Technology Commission of Shanghai Municipality[16DZ1910200]
WOS研究方向	Physiology
WOS类目	Physiology
WOS记录号	WOS:000471315000001
出版者	FRONTIERS MEDIA SA
WOS关键词	SIZE-EXCLUSION CHROMATOGRAPHY ; GAP-JUNCTION CHANNELS ; SKIN-DISEASE ; ATP RELEASE ; CELL-DEATH ; PANNEXIN 1 ; OCULODENTODIGITAL DYSPLASIA ; THERAPEUTIC ANTIBODIES ; CX26 HEMICHANNELS ; MUTATIONS
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/48971
专题	免疫化学研究所_公共科研平台_高通量筛选平台免疫化学研究所_特聘教授组_功能筛选实验室免疫化学研究所_特聘教授组_抗体设计学实验室免疫化学研究所_公共科研平台生命科学与技术学院_硕士生免疫化学研究所_特聘教授组_Michael Levitt组
通讯作者	Yang, Guang; Zonta, Francesco; Mammano, Fabio
作者单位	1.CNR, Inst Cell Biol & Neurobiol, Monterotondo, Italy 2.Univ Cattolica Sacro Cuore, Inst Otolaryngol, Rome, Italy 3.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai, Peoples R China 4.Univ Padua, Dept Phys & Astron G Gelilei, Padua, Italy 5.Univ A Gemelli IRCCS, Fdn Policlin, Rome, Italy 6.Deutsch Krebsforsch Zentrum DKFZ, Div Funct Genome Anal, German Canc Res Ctr DKFZ, Heidelberg, Germany
通讯作者单位	免疫化学研究所
推荐引用方式 GB/T 7714	Ziraldo, Gaia,Buratto, Damiano,Kuang, Yuanyuan,et al. A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function[J]. FRONTIERS IN PHYSIOLOGY,2019,10.
APA	Ziraldo, Gaia.,Buratto, Damiano.,Kuang, Yuanyuan.,Xu, Liang.,Carrer, Andrea.,...&Mammano, Fabio.(2019).A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function.FRONTIERS IN PHYSIOLOGY,10.
MLA	Ziraldo, Gaia,et al."A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function".FRONTIERS IN PHYSIOLOGY 10(2019).