Differential coupling of M1 muscarinic and alpha7 nicotinic receptors to inhibition of pemphigus acantholysis.
The mechanisms mediating and regulating assembly and disassembly of intercellular junctions is a subject of intensive research. The IgG autoantibodies produced in patients with the immunoblistering skin disease pemphigus vulgaris (PV) can induce keratinocyte (KC) dyshesion (acantholysis) via mechanisms that involve signaling kinases targeting intercellular adhesion molecules, thus providing a useful model to study the physiologic regulation of KC cohesion. Previous studies showed that activation of Src and protein kinase C are the earliest events in the PV IgG-induced intracellular phosphorylation cascades and that cholinergic agonists are effective for treating patients with pemphigus. In this study, we sought to elucidate the molecular mechanisms allowing cholinergic agonists to inhibit PV IgG-induced acantholysis and phosphorylation of KC adhesion molecules. The extent of acantholysis in KC monolayers correlated closely with the degree of PV IgG-induced phosphorylation of p120- and beta-catenins, with classic isoforms of protein kinase C mediating serine phosphorylation of beta-catenin and Src-tyrosine phosphorylation of p120-catenin. The M(1) muscarinic agonist pilocarpine blocked phosphorylation of both catenins, which could be abolised by the M(1) antagonist MT7. The alpha7 nicotinic agonist AR-R17779 inhibited phosphorylation of P120-cateinin. The alpha7 antagonist methyllycaconitine abolished the effect of AR-R17779. Okadaic acid abrogated protective effects of agonists on phosphorylation of beta-catenin, and pervanadate, on that of p120-catenin. Stimulation of KCs with pilocarpine significantly (p < 0.05) elevated both serine/threonine and tyrosine phosphatase activities in KCs. AR-R17779 both stimulated tyrosine phosphatase and decreased PV IgG-induced Src activity. Methyllycaconitine released Src activity in intact KCs and caused acantholysis. Thus, downstream signaling from M(1) abolished PV IgG-dependent catenin phosphorylation due to activation of both serine/threonine and tyrosine phosphatases, whereas alpha7 action involved both activation of tyrosine phosphatase and inhibition of Src. These findings identified novel paradigm of regulation of signaling kinases associated with cholinergic receptors and provided mechanistic explanation of therapeutic activity of cholinomimetics in PV patients.
Chernyavsky
Alex I
AI
Department of Dermatology, University of California, Irvine, California 92697.
Arredondo
Juan
J
Department of Dermatology, University of California, Irvine, California 92697.
Piser
Timothy
T
AstraZeneca Pharmaceuticals, Wilmington, Delaware 19850.
Karlsson
Evert
E
Karolinska Institute, Lindsbergsgatan 11A, Uppsala 752 40, Sweden.
Grando
Sergei A
SA
Department of Dermatology, University of California, Irvine, California 92697. Electronic address: sgrando@uci.edu.
D000157:Aconitine / Q000031:analogs & derivatives / Q000494:pharmacology; D002448:Cell Adhesion; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D008954:Models, Biological; D019319:Okadaic Acid / Q000378:metabolism; D010392:Pemphigus / Q000378:metabolism*; D010766:Phosphorylation; D043643:Receptor, Muscarinic M1 / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D064569:alpha7 Nicotinic Acetylcholine Receptor; D051176:beta Catenin / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
C579167:Chrna7 protein, human; D007074:Immunoglobulin G; D043643:Receptor, Muscarinic M1; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D051176:beta Catenin; D019319:Okadaic Acid; C054634:methyllycaconitine; D019061:src-Family Kinases; D000157:Aconitine