title: Carnosine metabolism in diabetes is altered by reactive metabolites.
abstract: Carnosinase 1 (CN1) contributes to diabetic nephropathy by cleaving histidine-dipeptides which scavenge reactive oxygen and carbonyl species and increase nitric oxide (NO) production. In diabetic mice renal CN1 activity is increased, the regulatory mechanisms are unknown. We therefore analysed the in vitro and in vivo regulation of CN1 activity using recombinant and human CN1, and the db/db mouse model of diabetes. Glucose, leptin and insulin did not modify recombinant and human CN1 activity in vitro, glucose did not alter renal CN1 activity of WT or db/db mice ex vivo. Reactive metabolite methylglyoxal and Fenton reagent carbonylated recombinant CN1 and doubled CN1 efficiency. NO S-nitrosylated CN1 and decreased CN1 efficiency for carnosine by 70 % (p < 0.01), but not for anserine. Both CN1 cysteine residues were nitrosylated, the cysteine at position 102 but not at position 229 regulated CN1 activities. In db/db mice, renal CN1 mRNA and protein levels were similar as in non-diabetic controls, CN1 efficiency 1.9 and 1.6 fold higher for carnosine and anserine. Renal carbonyl stress was strongly increased and NO production halved, CN1 highly carbonylated and less S-nitrosylated compared to WT mice. GSH and NO2/3 concentrations were reduced and inversely related with carnosine degradation rate (r = -0.82/-0.85). Thus, reactive metabolites of diabetes upregulate CN1 activity by post-translational modifications, and thus decrease the availability of reactive metabolite-scavenging histidine dipeptides in the kidney in a positive feedback loop. Interference with this vicious circle may represent a new therapeutic target for mitigation of DN.
authors.affiliation: Centre for Paediatric and Adolescence Medicine, University of Heidelberg, Heidelberg, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Peters
authors.affiliation: Division of Human Genetics, Medical University Innsbruck, Innsbruck, Austria.
authors.forename: Barbara
authors.initials: B
authors.lastname: Lanthaler
authors.affiliation: Division of Human Genetics, Medical University Innsbruck, Innsbruck, Austria.
authors.forename: Albert
authors.initials: A
authors.lastname: Amberger
authors.affiliation: Internal Medicine, University Heidelberg, Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Fleming
authors.affiliation: Department of Endocrinology, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Elisabete
authors.initials: E
authors.lastname: Forsberg
authors.affiliation: Institute for Physiology and Pathophysiology, University Heidelberg, Heidelberg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Hecker
authors.affiliation: Institute for Physiology and Pathophysiology, University Heidelberg, Heidelberg, Germany.
authors.forename: Andreas H
authors.initials: AH
authors.lastname: Wagner
authors.affiliation: Structural Genomics Consortium, University of Oxford, Oxford, UK.
authors.forename: Wyatt W
authors.initials: WW
authors.lastname: Yue
authors.affiliation: Centre for Paediatric and Adolescence Medicine, University of Heidelberg, Heidelberg, Germany.
authors.forename: Georg F
authors.initials: GF
authors.lastname: Hoffmann
authors.affiliation: Internal Medicine, University Heidelberg, Heidelberg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Nawroth
authors.affiliation: Division of Human Genetics, Medical University Innsbruck, Innsbruck, Austria. johannes.zschocke@i-med.ac.at.
authors.forename: Johannes
authors.initials: J
authors.lastname: Zschocke
authors.affiliation: Centre for Paediatric and Adolescence Medicine, University of Heidelberg, Heidelberg, Germany.
authors.forename: Claus P
authors.initials: CP
authors.lastname: Schmitt
mesh_terms: D000818:Animals; D002336:Carnosine / Q000235:genetics / Q000378:metabolism*; D003920:Diabetes Mellitus / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004150:Dipeptidases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains; D009154:Mutation; D009569:Nitric Oxide / Q000378:metabolism*; D011765:Pyruvaldehyde / Q000378:metabolism*
chemical_list: C045076:Fenton's reagent; D009569:Nitric Oxide; D011765:Pyruvaldehyde; D002336:Carnosine; D006861:Hydrogen Peroxide; D007501:Iron; D004150:Dipeptidases; C019673:aminoacyl-histidine dipeptidase
keywords: Anserine; Carnosinase; Carnosine; Diabetic nephropathy; N-carbonylation; S-nitrosylation