Hakim Boukhalfa

Chemical aspects of siderophore mediated iron transport

In this mini-review we describe selected aspects of the coordination chemistry relevant to siderophore mediated iron transport and bioavailability. Specific emphasis is placed on a discussion of in vitro kinetic and thermodynamic data that are relevant to elucidating possible in vivo mechanisms for environmental iron acquisition by microbial cells.

Plutonium(V/VI) Reduction by the Metal-Reducing Bacteria Geobacter metallireducens GS-15 and Shewanella oneidensis MR-1.

ABSTRACT We examined the ability of the metal-reducing bacteria Geobacter metallireducens GS-15 and Shewanella oneidensis MR-1 to reduce Pu(VI) and Pu(V). Cell suspensions of both bacteria reduced oxidized Pu [a mixture of Pu(VI) and Pu(V)] to Pu(IV). The rate of plutonium reduction was similar to the rate of U(VI) reduction obtained under similar conditions for each bacteria. The rates of Pu(VI) and U(VI) reduction by cell suspensions of S. oneidensis were slightly higher than the rates observed with G. metallireducens. The reduced form of Pu was characterized as aggregates of nanoparticulates of Pu(IV). Transmission electron microscopy images of the solids obtained from the cultures after the reduction of Pu(VI) and Pu(V) by S. oneidensis show that the Pu precipitates have a crystalline structure. The nanoparticulates of Pu(IV) were precipitated on the surface of or within the cell walls of the bacteria. The production of Pu(III) was not observed, which indicates that Pu(IV) was the stable form of reduced Pu under these experimental conditions. Experiments examining the ability of these bacteria to use Pu(VI) as a terminal electron acceptor for growth were inconclusive. A slight increase in cell density was observed for both G. metallireducens and S. oneidensis when Pu(VI) was provided as the sole electron acceptor; however, Pu(VI) concentrations decreased similarly in both the experimental and control cultures.

Kinetics and mechanism of iron release from the bacterial ferric binding protein nFbp: exogenous anion influence and comparison with mammalian transferrin

Ferric binding protein, Fbp, serves an essential biological function in shuttling naked (hydrated) Fe3+ across the periplasmic space of many Gram-negative bacteria. In this process, iron must be released at the cytoplasmic membrane to a permease. How iron is released from Fbp has yet to be resolved. Consequently, understanding the dynamics of iron release from Fbp is of both biological and chemical interest. Fbp requires an exogenous anion, e.g. phosphate when isolated from cell lysates, for tight iron sequestration. To address the role of exogenous anion identity and lability on Feaq3+ dissociation from Fbp, the kinetics of PO43− exchange in Fe3+nFbp(PO4) (nFbp=recombinant Fbp from Neisseria meningitidis) were investigated by dynamic 31P NMR and the kinetics of Fe3+ dissociation from Fe3+nFbp(X) (X=PO43−, citrate anion) were investigated by stopped-flow pH-jump measurements. We justify the use of non-physiological low-pH conditions because a high [H+] will drive the Feaq3+ dissociation reaction to completion without using competing chelators, whose presence may complicate or influence the dissociation mechanism. For perspective, these studies of nFbp (which has been referred to as a bacterial transferrin) are compared to new and previously published kinetic and thermodynamic data for mammalian transferrin. Significantly, we address the lability of the Fe3+ coordination shell in nFbp, Fe3+nFbp(X) (X=PO43−, citrate), with respect to exogenous anion (Xn−) exchange and dissociation, and ultimately complete dissociation of the protein to yield naked (hydrated) Feaq3+. These findings are a first step in understanding the process of iron donation to the bacterial permease for transport across the cytoplasmic membrane.

Beryllium(II) binding to ATP and ADP: Potentiometric determination of the thermodynamic constants and implications for in vivo toxicity

Highly toxic beryllium(II) is divalent metal ion with a high charge density, making it a potential target for binding to bio-molecules rich in O donor groups. In aqueous solution Be2+ binds to ATP and ADP to form 1:1 Be2+:ATP and Be2+:ADP complexes in relatively acidic media. At neutral pH the complex formed undergoes hydrolysis. Be2+ binding to ATP and ADP is much stronger than Ca2+ and Mg2+ binding. The high affinity of Be2+ toward ATP and ADP binding suggests a mechanism relevant to understanding the in vivo chemical toxicity of this metal.