Pannexins (Panx1, 2, 3) are channel-forming glycoproteins expressed in mammalian 
tissues. We previously reported that N-glycosylation acts as a regulator of the 
localization and intermixing of Panx1 and Panx3, but its effects on Panx2 are 
currently unknown. Panx1 and Panx2 intermixing can regulate channel properties, 
and both pannexins have been implicated in neuronal cell death after ischemia. 
Our objectives were to validate the predicted N-glycosylation site of Panx2 and 
to study the effects of Panx2 glycosylation on localization and its capacity to 
interact with Panx1. We used site-directed mutagenesis, enzymatic 
de-glycosylation, cell-surface biotinylation, co-immunoprecipitation, and 
confocal microscopy. Our results showed that N86 is the only N-glycosylation 
site of Panx2. Panx2 and the N86Q mutant are predominantly localized to the 
endoplasmic reticulum (ER) and cis-Golgi matrix with limited cell surface 
localization was seen only in the presence of Panx1. The Panx2 N86Q mutant is 
glycosylation-deficient and tends to aggregate in the ER reducing its cell 
surface trafficking but it can still interact with Panx1. Our study indicates 
that N-glycosylation may be important for folding and trafficking of Panx2. We 
found that the un-glycosylated forms of Panx1 and 2 can readily interact, 
regulating their localization and potentially their channel function in cells 
where they are co-expressed.