NMN deamidase (PncC) is a bacterial enzyme involved in NAD biosynthesis. We have previously demonstrated that PncC is structurally distinct from other known amidohydrolases. Here, we extended PncC characterization by mutating all potential catalytic residues and assessing their individual roles in catalysis through kinetic analyses. Inspection of these residues’ spatial arrangement in the active site, allowed us to conclude that PncC is a serine-amidohydrolase, employing a Ser/Lys dyad for catalysis. Analysis of the PncC structure in complex with a modeled NMN substrate supported our conclusion, and enabled us to propose the catalytic mechanism
Characterization of bacterial NMN deamidase as a Ser/Lys hydrolase expands diversity of serine amidohydrolases / Sorci, Leonardo; Brunetti, L; Cialabrini, L; Mazzola, Francesca; Kazanov, Md; D'Auria, S; Ruggieri, Silverio; Raffaelli, Nadia. - In: FEBS LETTERS. - ISSN 0014-5793. - 588:6(2014), pp. 1016-1023. [10.1016/j.febslet.2014.01.063]
Characterization of bacterial NMN deamidase as a Ser/Lys hydrolase expands diversity of serine amidohydrolases.
SORCI, Leonardo;MAZZOLA, FRANCESCA;RUGGIERI, Silverio;RAFFAELLI, Nadia
2014-01-01
Abstract
NMN deamidase (PncC) is a bacterial enzyme involved in NAD biosynthesis. We have previously demonstrated that PncC is structurally distinct from other known amidohydrolases. Here, we extended PncC characterization by mutating all potential catalytic residues and assessing their individual roles in catalysis through kinetic analyses. Inspection of these residues’ spatial arrangement in the active site, allowed us to conclude that PncC is a serine-amidohydrolase, employing a Ser/Lys dyad for catalysis. Analysis of the PncC structure in complex with a modeled NMN substrate supported our conclusion, and enabled us to propose the catalytic mechanismI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
