Naslov
Kinetics and Mechanism of Iron(III) Complexation by Ferric Binding Protein: The Role of Phosphate

Autori
Gabričević, Mario ; Anderson, Damon S ; Mietzner, Timothy A ; Crumbliss, Alvin L.

Izvornik
Biochemistry (0006-2960) 43 (2004), 19; 5811-5819

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Bacterial transferrin; ferric binding protein; iron transport; kinetics; mechanism; Neisseria meningitidis

Sažetak
Iron transport across the periplasmic space to the cytoplasmic membrane of certain gram negative bacteria is mediated by ferric binding protein (Fbp). This requires Fe3+ loading of Fbp at the inner leaflet of the outer membrane. A synergistic anion is required for tight Fe3+ sequestration by Fbp. Although phosphate fills this role in the protein isolated from bacterial cell lysates, NTA can also satisfy this requirement in vitro. Here we report the kinetics and mechanism of Fe3+ loading of Fbp from Fe(NTA)aq in the presence of phosphate. The reaction proceeds in four kinetically distinguishable steps to produce Fe3+Fbp(PO4) as a final product. The first three steps exhibit half-lives ranging from ca 20 msec to 0.5 min, depending on concentrations and produces Fe3+Fbp(NTA) as an intermediate product of significant stability. The rates of the first two steps are accelerated with increasing phosphate concentration, while that of the third step is retarded by phosphate. Conversion of Fe3+Fbp(NTA) to Fe3+Fbp(PO4) in the fourth step is a slow process (half-life ~2 hrs) and is facilitated by free phosphate. A mechanism for the Fe3+ loading process is proposed in which the synergistic anions phosphate and NTA play key roles. These data suggest that not only is the synergistic anion required for tight Fe3+ sequestration by Fbp, but the synergistic anion also plays an important role in the process of inserting Fe3+ into the Fbp binding site.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

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