Fast dissociation of nitric oxide from ferrous Pseudomonas aeruginosa cd1 nitrite reductase: A novel outlook on the catalytic mechanism

Alessandro Arcovito, Serena Rinaldo, Maurizio Brunori, Francesca Cutruzzolà

Risultato della ricerca: Contributo in rivistaArticolo in rivistapeer review

40 Citazioni (Scopus)

Abstract

The heme-containing periplasmic nitrite reductase (cd1 NIR) is responsible for the production of nitric oxide (NO) in denitrifying bacterial species, among which are several animal and plant pathogens. Heme NIRs are homodimers, each subunit containing one covalently bound c-heme and one d 1-heme. The reduction of nitrite to NO involves binding of nitrite to the reduced protein at the level of d1-heme, followed by dehydration of nitrite to yield NO and release of the latter. The crucial ratelimiting step in the catalytic mechanism is thought to be the release of NO from the d 1-heme, which has been proposed, but never demonstrated experimentally, to occur when the iron is in the ferric form, given that the reduced NO-bound derivative was presumed to be very stable, as in other hemeproteins. We have measured for the first time the kinetics of NO binding and release from fully reduced cd1 NIR, using the enzyme from Pseudomonas aeruginosa and its site-directed mutant H369A. Quite unexpectedly, we found that NO dissociation from the reduced d1-heme is very rapid, several orders of magnitude faster than that measured for b-type heme containing reduced hemeproteins. Because the rate of NO dissociation from reduced cd1 NIR, measured in the present report, is faster than or comparable with the turnover number, contrary to expectations this event may well be on the catalytic cycle and not necessarily rate-limiting. This finding also provides a rationale for the presence in cd1 NIR of the peculiar d1-heme cofactor, which has probably evolved to ensure fast product dissociation
Lingua originaleEnglish
pagine (da-a)14761-14767
Numero di pagine7
RivistaTHE JOURNAL OF BIOLOGICAL CHEMISTRY
Stato di pubblicazionePubblicato - 2007

Keywords

  • nitrite reductase

Fingerprint Entra nei temi di ricerca di 'Fast dissociation of nitric oxide from ferrous Pseudomonas aeruginosa cd1 nitrite reductase: A novel outlook on the catalytic mechanism'. Insieme formano una fingerprint unica.

Cita questo