title: Hydrogen peroxide regulates the cholinergic signal in a concentration dependent manner.
abstract: The human epidermis holds the full capacity for autocrine synthesis, transport and degradation of acetylcholine as well as the muscarinic (m1-m5) and nicotinic signal transduction in keratinocytes and melanocytes. This cholinergic cascade is severely affected in patients with the depigmentation disorder vitiligo due to accumulation of hydrogen peroxide (H(2)O(2)) in the mM range as shown by in vivo FT-Raman spectroscopy. These high levels can oxidise susceptible amino acid residues such as methionine, tryptophan, cysteine and selenocysteine in the structure of proteins and peptides which in turn can severely affect the function. Here the effect of this reactive oxygen species was followed on the production and degradation of acetylcholine using immunofluorescence, enzyme kinetics, in vivo and in vitro FT-Raman and fluorescence spectroscopy as well as computer modelling. The results showed that both epidermal acetylcholinesterase (AchE) and butyrylcholinesterase (BchE) are target to H(2)O(2)-mediated oxidation of methionine and tryptophan residues close to the catalytic triad, while cholineacetyltransferase (chAT) is not affected. Enzyme kinetics revealed concentration dependent activation/deactivation of both degrading enzymes by H(2)O(2). Oxidation of methionine to methionine sulfoxide was confirmed by FT-Raman spectroscopy while oxidation of tryptophan to 5OH-tryptophan was identified by fluorescence spectroscopy. H(2)O(2)-mediated oxidation of both enzymes takes place in acute vitiligo yielding accumulation of acetylcholine in the epidermis of these patients. This process is reversible with a narrowband UVB activated pseudocatalase PC-KUS leading to recovery of epidermal and systemic enzyme activities as well as restoration of the lost skin colour.
authors.affiliation: Clinical and Experimental Dermatology/Department of Biomedical Sciences, University of Bradford, Bradford, BD7 1DP, UK. K.Schallreuter@bradford.ac.uk
authors.forename: Karin U
authors.initials: KU
authors.lastname: Schallreuter
authors.forename: Souna
authors.initials: S
authors.lastname: Elwary
mesh_terms: D000109:Acetylcholine / Q000737:chemistry / Q000378:metabolism*; D000110:Acetylcholinesterase / Q000737:chemistry / Q000378:metabolism*; D002091:Butyrylcholinesterase / Q000737:chemistry / Q000378:metabolism*; D002374:Catalase / Q000627:therapeutic use; D004305:Dose-Response Relationship, Drug; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology*; D008958:Models, Molecular; D014820:Vitiligo / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control
chemical_list: D006861:Hydrogen Peroxide; C038109:pseudocatalase; D002374:Catalase; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D000109:Acetylcholine