%0 Journal Article
%T Evaluation of the Phosphoproteome of Mouse Alpha 4/Beta 2-Containing Nicotinic Acetylcholine Receptors In Vitro and In Vivo
%A Angus C. Nairn
%A Marina R. Picciotto
%A Megan B. Miller
%A Rashaun S. Wilson
%A TuKiet T. Lam
%J -
%D 2018
%R https://doi.org/10.3390/proteomes6040042
%X Abstract Activation of nicotinic acetylcholine receptors containing ¦Á4 and ¦Â2 subunits (¦Á4/¦Â2* nAChRs) in the mammalian brain is necessary for nicotine reinforcement and addiction. We previously identified interactions between ¦Á4/¦Â2* nAChRs and calcium/calmodulin-dependent protein kinase II (CaMKII) in mouse and human brain tissue. Following co-expression of ¦Á4/¦Â2 nAChR subunits with CaMKII in HEK cells, mass spectrometry identified 8 phosphorylation sites in the ¦Á4 subunit. One of these sites and an additional site were identified when isolated ¦Á4/¦Â2* nAChRs were dephosphorylated and subsequently incubated with CaMKII in vitro, while 3 phosphorylation sites were identified following incubation with protein kinase A (PKA) in vitro. We then isolated native ¦Á4/¦Â2* nAChRs from mouse brain following acute or chronic exposure to nicotine. Two CaMKII sites identified in HEK cells were phosphorylated, and 1 PKA site was dephosphorylated following acute nicotine administration in vivo, whereas phosphorylation of the PKA site was increased back to baseline levels following repeated nicotine exposure. Significant changes in ¦Â2 nAChR subunit phosphorylation were not observed under these conditions, but 2 novel sites were identified on this subunit, 1 in HEK cells and 1 in vitro. These experiments identified putative CaMKII and PKA sites on ¦Á4/¦Â2* nAChRs and novel nicotine-induced phosphorylation sites in mouse brain that can be explored for their consequences on receptor function. View Full-Tex
%K nicotinic receptor
%K CaMKII
%K PKA
%K quantitative phosphoproteomics
%K mouse
%K phosphorylation
%K nicotine
%U https://www.mdpi.com/2227-7382/6/4/42