Browsing by Author "Edwards PJB"

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    Cell wall permeability in relation to in vitro starch digestion of pea cotyledon cells
    (Elsevier B.V., 2024-08-07) Ajala A; Kaur L; Lee SJ; Edwards PJB; Singh J
    The role of cell wall permeability and rate of starch digestibility in intact cotyledon cells from four different varieties of pea seeds was studied. Pulsed-field gradient nuclear magnetic resonance (PFG NMR) coupled with light and confocal microscopy were employed to evaluate the cotyledon cells' diffusion coefficients and cell wall permeability. The cotyledon cells' diffusion coefficients and cell wall permeability followed a decreasing trend: White/yellow pea > Marrowfat pea > Maple pea > Blue pea. The varying size of internal cavities in the microstructure in the cotyledon cells, as observed by the light and confocal micrographs, may be responsible for this trend. The extent of starch hydrolysis recorded from the cotyledon cells followed the same trend of the cell wall permeability except for Blue pea cotyledon cells. Thus, indicating that the more permeable the cotyledon cell to the starch-degrading enzymes, the higher the extent of intracellular starch hydrolysis. The microstructure changes in the cotyledon cells during digestion also confirmed this observation.
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    Does harvesting age matter? Changes in structure and rheology of a shear-thickening polysaccharide from Cyathea medullaris as a function of age
    (Elsevier Ltd, 2024-04-01) Bisht A; Goh KKT; Sims IM; Edwards PJB; Matia-Merino L
    A shear-thickening polysaccharide from the New Zealand Black tree fern (Cyathea medullaris, commonly known as mamaku) extracted from different age fronds (stage 1: young, stage 2: fully grown and stage 3: old) was characterised in terms of structure and rheological properties. Constituent sugar analysis and 1H and 13C NMR revealed a repeating backbone of −4)-β-D-GlcpA-(1 → 2)-α-D-Manp-(1→, for all mamaku polysaccharide (MP) samples from different age fronds without any alterations in molecular structure. However, the molecular weight (Mw) was reduced with increasing age, from ~4.1 × 106 to ~2.1 × 106 Da from stage 1 to stage 3, respectively. This decrease in Mw (and size) consequently reduced the shear viscosity (ηs-Stage 1 > ηs-Stage 2 > ηs-Stage 3). However, the extent of shear-thickening and uniaxial extensional viscosity of MP stage 2 was greater than MP stage 1, which was attributed to a greater intermolecular interaction occurring in the former. Shear-thickening behaviour was not observed in MP stage 3.
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    Inverted strand polarity yields thermodynamically stable G-quadruplexes and prevents duplex formation within extended DNA.
    (The Royal Society of Chemistry, 2024-08-27) Chilton B; Roach RJ; Edwards PJB; Jameson GB; Hale TK; Filichev VV
    DNA G-quadruplexes (G4) formed in guanine-rich sequences play a key role in genome function and maintenance, interacting with multiple proteins. However, structural and functional studies of G4s within duplex DNA have been challenging because of the transient nature of G4s and thermodynamic preference of G-rich DNA to form duplexes with their complementary strand rather than G4s. To overcome these challenges, we have incorporated native nucleotides in G-rich sequences using commercially available inverted 3'-O-DMT-5'-O-phosphoramidites of native nucleosides, to give 3'-3' and 5'-5' linkages in the centre of the G-tract. Using circular dichroism and 1H nuclear magnetic resonance spectroscopies and native gel electrophoresis, we demonstrate that these polarity-inverted DNA sequences containing four telomeric repeats form G4s of parallel topology with one lateral or diagonal loop across the face of the quadruplex and two propeller loops across the edges of the quadruplex. These G4s were stable even in the presence of complementary C-rich DNA. As an example, G4 assemblies of inverted polarity were shown to bind to the hinge region of Heterochromatin Protein 1α (HP1α), a known G4-interacting domain. As such, internal polarity inversions in DNA provide a useful tool to control G4 topology while also disrupting the formation of other secondary structures, particularly the canonical duplex.