| Budget Amount *help |
¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
|
|---|
| Research Abstract |
This study is aimed to investigate the mechanism of the biosynthesis of arabinogalactan-proteins (AGPs) involved in cell walls of higher plants, and to evaluate their physiological roles concerning to tissue differentiation in relation to the organ specific expression of their sugar chains. To approach this purpose, alpha-L-fucosyltransferse (alpha-L-FucTase), an enzyme catalyzing the transfer of L-fucose (L-Fuc) residues, is focused based on its specific assay for enzyme activity and enzymatic properties, in addition to the localization of the enzyme in sub cellular organs.
1.The reaction mixture was constructed by GDP-L-Fuc as the sugar donor and and Araalpha1*3Galbeta1*6Gal derivatized with 2-aminopyridine (AGG-Pa) as the acceptor substrate together with the microsome (crude membrane) fraction as the enzyme source obtained from 6-d-old radish primary roots. After incubation at 25C,the enzyme activity was assayd by estimation of the transferred product separated by HPLC.The enzyme assay EAS also done by using GDP-L- [^<14>C] -Fuc in order to estimate the tranfer action into high molecular weight compounds.
2.The enzyme properties : The enzyme is maximally active at pH6.0-7.5 and at 25oC,and requires Zwittergent and Mn^<21> or Mg^2. The km values are estimated to be 0.08 and 3.72mM for GDP-L-Fuc and AGG-PA,respectively. Chemical and enzymatic analyzes of fucosylated AGG-PA confirmed the attachment of L-Fuc to the arabinosyl residue at 0-2 by alpha-glycosidic linkage.
|
