The Liver Connexin32 Interactome Is a Novel Plasma Membrane-Mitochondrial Signaling Nexus

doi:10.1021/pr301166p

	The Liver Connexin32 Interactome Is a Novel Plasma Membrane-Mitochondrial Signaling Nexus
	Fowler, Stephanie L.1,3; Akins, Mark 1,3; Zhou, Hu2 ; Figeys, Daniel ; Bennett, Steffany A. L.1,3
刊名	JOURNAL OF PROTEOME RESEARCH
	2013-06
卷号	12 期号:6 页码:2597-2610
关键词	gap junction connexin connexin32 connexin26 mitochondria interactome sideroflexin mass spectrometry immunoprecipitation inner mitochondrial membrane
ISSN号	1535-3893
DOI	10.1021/pr301166p
文献子类	Article
英文摘要	Connexins are the structural subunits of gap junctions and act as protein platforms for signaling complexes. Little is known about tissue-specific connexin signaling nexuses, given significant challenges associated with affinity-purifying endogenous channel complexes to the level required for interaction analyses. Here, we used multiple subcellular fractionation techniques to isolate connexin32-enriched membrane microdomains from murine liver. We show, for the first time, that connexin32 localizes to both the plasma membrane and inner mitochondrial membrane of hepatocytes. Using a combination of immunoprecipitation-high throughput mass spectrometry, reciprocal co-IP, and subcellular fractionation methodologies, we report a novel interactome validated using null mutant controls. Eighteen connexin32 interacting proteins were identified. The majority represent resident mitochondrial proteins, a minority represent plasma membrane, endoplasmic reticulum, or cytoplasmic partners. In particular, connexin32 interacts with connexin26 and the mitochondrial protein, sideroflexin-1, at the plasma membrane. Connexin32 interaction enhances connexin26 stability. Converging bioinformatic, biochemical, and confocal analyses support a role for connexin32 in transiently tethering mitochondria to connexin32-enriched plasma membrane microdomains through interaction with proteins in the outer mitochondrial membrane, including sideroflexin-1. Complex formation increases the pool of sideroflexin-1 that is present at the plasma membrane. Together, these data identify a novel plasma membrane/mitochondrial signaling nexus in the connexin32 interactome.
资助项目	CIHR[MOP 62826] ; CIHR Training Program in Neurodegenerative Lipidomics (CTPNL)[TGF 96121] ; Institute of Aging[00000000] ; CTPNL[00000000]
WOS关键词	ADENINE-NUCLEOTIDE TRANSLOCATOR ; JUNCTIONAL INTERCELLULAR COMMUNICATION ; GAP-JUNCTIONS ; SIDEROFLEXIN FAMILY ; RAT HEPATOCYTES ; PROTEINS ; MOUSE ; DOMAINS ; GENES ; EXPRESSION
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000320298600023
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/277608]
专题	分析化学研究室
通讯作者	Figeys, Daniel
作者单位	1.Univ Ottawa, Neural Regenerat Lab, Ottawa, ON, Canada; 2.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 200031, Peoples R China 3.Univ Ottawa, Ottawa Inst Syst Biol, Dept Biochem Microbiol & Immunol, Ottawa, ON, Canada;
推荐引用方式 GB/T 7714	Fowler, Stephanie L.,Akins, Mark,Zhou, Hu,et al. The Liver Connexin32 Interactome Is a Novel Plasma Membrane-Mitochondrial Signaling Nexus[J]. JOURNAL OF PROTEOME RESEARCH,2013,12(6):2597-2610.
APA	Fowler, Stephanie L.,Akins, Mark,Zhou, Hu,Figeys, Daniel,&Bennett, Steffany A. L..(2013).The Liver Connexin32 Interactome Is a Novel Plasma Membrane-Mitochondrial Signaling Nexus.JOURNAL OF PROTEOME RESEARCH,12(6),2597-2610.
MLA	Fowler, Stephanie L.,et al."The Liver Connexin32 Interactome Is a Novel Plasma Membrane-Mitochondrial Signaling Nexus".JOURNAL OF PROTEOME RESEARCH 12.6(2013):2597-2610.