Biomedical subjects

E A Venczel

Publications and source records attributed to E A Venczel.

3 recordsLinked to original sources

PubMed1998

Activation of CTP:phosphocholine cytidylyltransferase by hypochlorite-oxidized phosphatidylcholines.

CTP:phosphocholine cytidylyltransferase (CT) catalyzes a rate-limiting, regulatory step in mammalian biosynthesis of phosphocholine (PC). Anionic phospholipids, fatty acids and diacylglycerol activate CT and promote its intercalation into the lipid bilayer, whereas zwitterionic phospholipids such as phosphatidylcholines do not. We investigated the effectiveness of polyunsaturated phosphatidylcholines as CT activators after hypochlorite oxidation. Detection and quantitation of oxidized PCs were evaluated by thin layer chromatography, high performance liquid chromatography, and conjugated dienes. Purified CT was assayed in the presence of multilamellar vesicles, containing variable concentrations of oxidized and parent PCs. The results demonstrate that particular species of oxidized PCs activate CT as potently as anionic lipids. The greater the number of double bonds available for oxidation in the fatty acid at the sn-2 position of the PC, the more effective was the oxidized PC as an activator of CT. Oxidized phospholipids at 1:1 bleach/lipid activated CT in the following order: PAPC>PL3PC>PL2PC compared to unoxidized controls. Since oxidized phospholipids decrease bilayer order (M.L. Wratten et al., Biochemistry 31 (1992) 10901-10907) these results are consistent with the activation of CT by perturbations of lipid bilayer packing.

Enzyme Activation↗

PubMed1996

Synapsable DNA.

We describe a simple innovation that allows DNA double helices to stably bind one another at specific sites, with regulatable affinity, under physiological conditions. This type of DNA synapsis requires neither an unraveling of the participating duplexes not heteroduplex formation, and is achieved by the intermolecular dimerization of short blocks of guanine-guanine mismatch base-pairs introduced within standard Watson-Crick duplexes. We propose that in vivo such "sticky" guanine domains, formed transiently in cruciforms, could initiate illegitimate recombination events. In practical terms, this type of synapsis, achievable in vitro by simply mixing the participating duplexes, could provide a novel and general technology for the self-assembly of arrays of important DNA sequences, and serve as a tool for investigating certain protein-DNA interactions in vivo.

Base Sequence↗

PubMed1993

Parallel and antiparallel G-DNA structures from a complex telomeric sequence.

We investigated the formation in vitro of higher order structures by a DNA oligomer containing the terminal motif TGTG3TGTGTGTG3, derived from the Saccharomyces telomeric consensus, in order to (a) understand why certain cations favor the formation of parallel-stranded (G4 and G8) G-DNA structures, while others favor foldback, antiparallel structures (G'2) and (b) probe the structures of G-DNAs formed by this telomeric sequence, which is more complex than its well-studied counterparts from the protozoans oxytricha and tetrahymena. We find that dramatic switches in the formation of G4 versus G'2 structures occur in solutions of not only the group Ia cations, Li(+)-Cs+, but also in those of the group IIa cations, Mg(2+)-Ba2+. These data and the temperature-dependent formation and destruction of the different structures lend support to the kinetic scheme of Sen and Gilbert (1990), by which rapidly forming G'2 structures accumulate in highly stabilizing potassium (and strontium) solutions at the expense of the thermodynamically more stable G4 structures. Both the G4 and the G'2 complexes formed by the Saccharomyces sequence show novel structural features. Protection and interference experiments with dimethyl sulfate and potassium permanganate reveal that the core of alternating thymines and guanines within the telomeric motif plays a critical role in the stabilization of the parallel G4 structure, but not of the antiparallel G'2. Very likely, in the G4 complex, this GT core forms a novel higher order arrangement of alternating G and T quartets, the latter possibly comparable to the U quartets described by Cheong and Moore (1992) in their NMR study of the higher order structure formed by rUG4U.

Base Sequence↗