Observation of a partially opened triple-helix conformation in 1-->3-beta-glucan by fluorescence resonance energy transfer spectroscopy.
This study used fluorescence resonance energy transfer (FRET) spectroscopy as an indirect method to investigate the effect of NaOH treatment on the conformation of a triple-helix (1-->3)-beta-D-glucan and then evaluated the effect of conformation on biological activity. Previous studies have suggested that treatment of the triple-helix glucans with NaOH produces single-helix conformers. FRET spectra of the triple-helix glucan, laminarin, doubly labeled with 1-aminopyrene as donor probe and fluorescein-5-isothiocyanate as acceptor probe attached at the reducing end, showed that a partially opened triple-helix conformer was formed on treatment with NaOH. Increasing degrees of strand opening was associated with increasing concentrations of NaOH. Based on these observations we propose that a partially opened triple-helix rather than a single helix, is formed by treating the triple-helix glucans with NaOH. After neutralizing the NaOH, changes in FRET indicated that the partially opened conformer gradually reverts to the triple-helix over 8 days. Laminarian was stabilized at different degrees of partial opening and its biological activity examined using the Limulus amebocyte lysate assay and nitric oxide production by alveolar macrophage. Both Limulus amebocyte lysate activity and nitric oxide production were related to the degree of opening of the triple-helix. Partially open conformers were more biologically active than the intact triple-helix.
Young
S H
SH
Department of Environmental Health Sciences, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Dong
W J
WJ
Jacobs
R R
RR
D000814:Aniline Compounds; D000818:Animals; D002236:Carbohydrate Conformation; D004735:Energy Transfer; D005456:Fluorescent Dyes; D005936:Glucans / Q000737:chemistry*; D006737:Horseshoe Crabs; D016676:Macrophages, Alveolar / Q000378:metabolism; D008970:Molecular Weight; D009569:Nitric Oxide / Q000378:metabolism; D012972:Sodium Hydroxide / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D047071:beta-Glucans
D000814:Aniline Compounds; D005456:Fluorescent Dyes; D005936:Glucans; D047071:beta-Glucans; D009569:Nitric Oxide; D012972:Sodium Hydroxide; C017006:aniline blue; C033363:beta-1,3-glucan