In humans, low serum carnosinase (CN1) activity protects patients with type 2 diabetes from diabetic nephropathy. We now characterized the interaction of thiol-containing compounds with CN1 cysteine residue at position 102, which is important for CN1 activity. Reduced glutathione (GSH), N-acetylcysteine and cysteine (3.2 Â± 0.4, 2.0 Â± 0.3, 1.6 Â± 0.2 Âµmol/mg/h/mM; p <.05) lowered dose-dependently recombinant CN1 (rCN1) efficiency (5.2 Â± 0.2 Âµmol/mg/h/mM) and normalized increased CN1 activity renal tissue samples of diabetic mice. Inhibition was allosteric. Substitution of rCN1 cysteine residues at position 102 (Mut1C102S) and 229 (Mut2C229S) revealed that only cysteine-102 is influenced by cysteinylation. Molecular dynamic simulation confirmed a conformational rearrangement of negatively charged residues surrounding the zinc ions causing a partial shift of the carnosine ammonium head and resulting in a less effective pose of the substrate within the catalytic cavity and decreased activity. Cysteine-compounds influence the dynamic behaviour of CN1 and therefore present a promising option for the treatment of diabetes.
Allosteric inhibition of carnosinase (CN1) by inducing a conformational shift / V. Peters, C.P. Schmitt, T. Weigand, K. Klingbeil, C. Thiel, A. van den Berg, V. Calabrese, P. Nawroth, T. Fleming, E. Forsberg, A.H. Wagner, M. Hecker, G. Vistoli. - In: JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY. - ISSN 1475-6366. - 32:1(2017), pp. 1102-1110.
|Titolo:||Allosteric inhibition of carnosinase (CN1) by inducing a conformational shift|
VISTOLI, GIULIO (Ultimo)
|Parole Chiave:||allosteric inhibition; Carnosinase 1 activity; CN1; diabetes; glutathione; N-acetylcysteine; allosteric regulation; dipeptidases; enzyme inhibitors; humans; molecular conformation; molecular dynamics simulation; sulfhydryl compounds; pharmacology; drug discovery; pharmaceutical science|
|Settore Scientifico Disciplinare:||Settore CHIM/08 - Chimica Farmaceutica|
|Data di pubblicazione:||2017|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1080/14756366.2017.1355793|
|Appare nelle tipologie:||01 - Articolo su periodico|