Glycosylation regulates pannexin intermixing and cellular localization.
The pannexin family of mammalian proteins, composed of Panx1, Panx2, and Panx3, has been postulated to be a new class of single-membrane channels with functional similarities to connexin gap junction proteins. In this study, immunolabeling and coimmunoprecipitation assays revealed that Panx1 can interact with Panx2 and to a lesser extent, with Panx3 in a glycosylation-dependent manner. Panx2 strongly interacts with the core and high-mannose species of Panx1 but not with Panx3. Biotinylation and dye uptake assays indicated that all three pannexins, as well as the N-glycosylation-defective mutants of Panx1 and Panx3, can traffic to the cell surface and form functional single-membrane channels. Interestingly, Panx2, which is also a glycoprotein and seems to only be glycosylated to a high-mannose form, is more abundant in intracellular compartments, except when coexpressed with Panx1, when its cell surface distribution increases by twofold. Functional assays indicated that the combination of Panx1 and Panx2 results in compromised channel function, whereas coexpressing Panx1 and Panx3 does not affect the incidence of dye uptake in 293T cells. Collectively, these results reveal that the functional state and cellular distribution of mouse pannexins are regulated by their glycosylation status and interactions among pannexin family members.
Penuela
Silvia
S
Department of Anatomy and Cell Biology, University of Western Ontario, London, Ontario N6A 5C1, Canada.
Bhalla
Ruchi
R
Nag
Kakon
K
Laird
Dale W
DW
D000818:Animals; D019921:Biotinylation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004396:Coloring Agents / Q000378:metabolism; D017630:Connexins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003911:Dextrans / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007473:Ion Channels / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000378:metabolism
D004396:Coloring Agents; D017630:Connexins; D003911:Dextrans; D007473:Ion Channels; D009419:Nerve Tissue Proteins; C503631:Panx1 protein, mouse; C516139:Panx2 protein, mouse; D011993:Recombinant Fusion Proteins; D012235:Rhodamines; C528477:pannexin 3 protein, mouse; C051311:rhodamine dextran; D008358:Mannose
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