Steady state and time-resolved fluorescence studies of a hemagglutinin from Moringa oleifera

TitleSteady state and time-resolved fluorescence studies of a hemagglutinin from Moringa oleifera
Publication TypeJournal Article
Year of Publication2008
AuthorsKatre, UV, Suresh, CG, Khan, MIslam, Gaikwad, SM
JournalJournal of Fluorescence
Volume18
Issue2
Pagination479-485
Date PublishedMAR
ISSN1053-0509
KeywordsANS binding, fluorescence, Hemagglutinin, lifetime, Moringa oleifera, saccharide binding, Solute quenching
Abstract

The saccharide binding and conformational characterization of a hemagglutinin, a low molecular weight protein from the seeds of Moringa oleifera was studied using steady state and time resolved fluorescence. The lectin binds sugars LacNAc (K-a = 1380 M-1) and fructose (K-a = 975 M-1), as determined by the fluorescence spectroscopy. It has a single tryptophan per monomer which is exposed on the surface and is in a strong electropositive environment as revealed by quenching with iodide. Quenching of the fluorescence by acrylamide involved both static (K-s = 0.216 M-1) and collisional (K-sv= 8.19 M-1) components. The native protein showed two different lifetimes, tau(1) (1.6 ns) and tau(2) (4.36 ns) which decrease and get converted into a single one, (2.21 ns) after quenching with 0.15 M acrylamide. The bimolecular quenching constant, k(q) was 7.55 x 10(11) M-1 s(-1). ANS binding studies showed that the native protein has exposed hydrophobic patches which get further exposed at extreme acidic or alkaline pH. However, they get buried in the interior of the protein in presence of 1 M GdnHCl or urea.

DOI10.1007/s10895-007-0289-7
Type of Journal (Indian or Foreign)Foreign
Impact Factor (IF)1.966
Divison category: 
Biochemical Sciences