Renalase is a recently discovered flavoprotein that regulates blood pressure, regulates sodium and phosphate excretion, and displays cardioprotectant action through a mechanism that is barely understood to date. It has been proposed to act as a catecholamine-degrading enzyme, via either O2-dependent or NADH-dependent mechanisms. Here we report the renalase crystal structure at 2.5 Å resolution together with new data on its interaction with nicotinamide dinucleotides. Renalase adopts the p-hydroxybenzoate hydroxylase fold topology, comprising a Rossmann-fold-based flavin adenine dinucleotide (FAD)-binding domain and a putative substrate-binding domain, the latter of which contains a five-stranded anti-parallel β-sheet. A large cavity (228 Å3), facing the flavin ring, presumably represents the active site. Compared to monoamine oxidase or polyamine oxidase, the renalase active site is fully solvent exposed and lacks an 'aromatic cage' for binding the substrate amino group. Renalase has an extremely low diaphorase activity, displaying lower kcat but higher kcat/Km for NADH compared to NADPH. Moreover, its FAD prosthetic group becomes slowly reduced when it is incubated with NADPH under anaerobiosis, and binds NAD+ or NADP+ with K d values of ca 2 mM. The absence of a recognizable NADP-binding site in the protein structure and its poor affinity for, and poor reactivity towards, NADH and NADPH suggest that these are not physiological ligands of renalase. Although our study does not answer the question on the catalytic activity of renalase, it provides a firm framework for testing hypotheses on the molecular mechanism of its action.
FAD-binding site and NADP reactivity in human renalase: A new enzyme involved in blood pressure regulation / M. Milani, F. Ciriello, S. Baroni, V. Pandini, G. Canevari, M. Bolognesi, A. Aliverti. - In: JOURNAL OF MOLECULAR BIOLOGY. - ISSN 0022-2836. - 411:2(2011), pp. 463-473.
|Titolo:||FAD-binding site and NADP reactivity in human renalase: A new enzyme involved in blood pressure regulation|
CIRIELLO, FRANCESCO (Secondo)
BOLOGNESI, MARTINO (Penultimo)
ALIVERTI, ALESSANDRO (Ultimo)
|Parole Chiave:||chronic kidney disease; end-stage renal disease; hypertension; phosphate excretion; flavin nucleotide; flavoprotein; enzyme; protein structure|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore BIO/11 - Biologia Molecolare
Settore BIO/09 - Fisiologia
|Data di pubblicazione:||2011|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.jmb.2011.06.010|
|Appare nelle tipologie:||01 - Articolo su periodico|
File in questo prodotto:
|JMB-D-11-00444R2 Author postprint.pdf||Articolo completo||Post-print, accepted manuscript ecc. (versione accettata dall'editore)||Administrator Richiedi una copia|
|Renalase crystal structure Milani Baroni Ciriello 2011_JMolBiol.pdf||Publisher's version/PDF||Administrator Richiedi una copia|