Synthesis, structure, and molecular dynamics of gallium complexes of schizokinen and the amphiphilic siderophore acinetoferrin

Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA.
Journal of the American Chemical Society (Impact Factor: 11.44). 10/2004; 126(38):12065-75. DOI: 10.1021/ja048145j
Source: PubMed

ABSTRACT A new general synthesis of the citrate-based siderophores acinetoferrin (Af) and schizokinen (Sz) and their analogues is described. The molecular structure of gallium schizokinen, GaSz, was determined by combined (1)H NMR, Hartree-Fock ab initio calculations, DFT, and empirical modeling of vicinal proton NMR spin-spin couplings. The metal-coordination geometry of GaSz was determined from NOE contacts to be cis-cis with respect to the two chelating hydroxamates. One diaminopropane adopts a single chairlike conformation while another is a mixture of two ring pucker arrangements. Both amide hydrogens are internally hydrogen bonded to metal-ligating oxygen atoms. The acyl methyl groups are directed away from each other with an average planar angle of ca. 130 degrees. The kinetics of GaSz racemization were followed by selective, double spin-echo inversion-recovery (1)H NMR spectroscopy over the temperature range of 10-45 degrees C. The racemization proceeds by a multistep mechanism that is proton independent between pD 5 and 12 (k(0) = 1.47 (0.15 s(-1))) and acid catalyzed below pD 4 (k(1) = 2.25 (0.15) x 10(4) M(-1) s(-1)). The activation parameters found for the two sequential steps of the proton independent pathway were DeltaH(++) = 25 +/- 3 kcal M(-1), DeltaS(++) = 25 +/- 7 cal M(-1) K(-1) and DeltaH(++) = 17.1 +/- 0.2 kcal M(-1), DeltaS(++) = 0.3 +/- 2.7 cal M(-1) K(-1). The first step of the proton-independent mechanism was assigned to the dissociation of the carboxyl group. The second step was assigned to complex racemization. The proton-assisted step was assigned to a complete dissociation of the alpha-hydroxy carboxyl group at pD < 4. The ab initio modeling of gallium acinetoferrin, GaAf, and analogues derived from the structure of GaSz has shown that the pendant trans-octenoyl fragments are oriented in opposite directions with the average planar angle of ca. 130 degrees. This arrangement prevents GaAf from adopting a phospholipid-like structural motif. Significantly, iron siderophore complex FeAf was found to be disruptive to phospholipid vesicles and is considerably more hydrophilic than Af, with an eight-fold smaller partition coefficient.

1 Follower
  • [Show abstract] [Hide abstract]
    ABSTRACT: The antimalarial drug chloroquine binds to gallium proto-porphyrin-IX in methanol and in the solid state and represents a unique drug-heme model.
    Chemical Communications 09/2014; 50(89). DOI:10.1039/C4CC05328A · 6.72 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Iron-limited cyanobacterial cells are generally considered to acquire extracellular iron through a siderophore-dependent system, although evidence has started to accumulate that other, as yet poorly characterized, iron acquisition systems may also play a role. Iron-limited cells of the cyanobacterium Anabaena flos-aquae (Lyng.) Brèb. are well known to produce the relatively low Fe(III) affinity dihydroxamate siderophore schizokinen. In this set of experiments we show that iron-limited A. flos-aquae cells (i) acquired iron at substantial rates in the absence of the schizokinen and (ii) acquired iron from a bacterial siderophore (the trihydroxamate molecule desferrioxamine B (DFB)), and also a synthetic chelator (N, N-bis(2-(bis(carboxymethyl)amino)ethyl)glycine (DTPA)), with substantially higher affinities for Fe(III) than schizokinen, indicating that a schizokinen-independent iron acquisition pathway was operating. We suggest that there exists a siderophore-independent iron acquisition system that is able to acquire Fe(III) from high stability Fe(III)-chelates, which are not accessible to iron-limited cells via the schizokinen-based system. As well, we present two possible models for iron acquisition by iron-limited A. flos-aquae cells. Both of these models suggest that there are two major routes for Fe(III) entry into the periplasm of iron-limited cells: (1) the well-characterized siderophore (schizokinen) dependent process and (2) a siderophore-independent process that is able to access Fe(III) sources not biologically available to the schizokinen system.
    Botany 03/2012; 90(3):181-190. DOI:10.1139/b11-099 · 1.04 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Under iron-deficient conditions, the Gram-negative bacterium Pseudomonas aeruginosa ATCC 15692 secretes a peptidic siderophore, pyoverdin PaA, composed of an aromatic chromophore derived from 2,3-diamino-6,7-dihydroxyquinoline and a partially cyclized octapeptide, D-Ser-L-Arg-D-Ser-L-FoOHOrn-(L-Lys-L-FoOHOrn-L-Thr-L-Thr) (FoOHOrn: delta N-formyl-delta N-hydroxyornithine), in which the C-terminal carboxyl group forms a peptidic bond with the primary amine of the L-Lys side chain. Ferric iron is chelated by the catechol group on the chromophore and the two hydroxyornithine side chains. In aqueous solution, the (1)H-NMR spectrum of pyoverdin PaA-Ga(III), in which Ga(III) is used instead of Fe(III) for spectroscopic purposes, showed clear evidence of exchange broadening, preventing further structural characterization. The use of cryo-solvents allowed measurements to be made at temperatures as low as 253 K where two distinct conformations with roughly equivalent populations could be observed. (13)C and (15)N labeling of pyoverdin PaA enabled complete assignment of both forms of pyoverdin PaA-Ga(III) at 253 and 267 K, using triple-resonance multidimensional NMR experiments commonly applied to doubly labeled proteins.
    Biopolymers 10/2005; 79(3):139-49. DOI:10.1002/bip.20343 · 2.29 Impact Factor


Available from