| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
Glycophorin is a transmembrane sialoglycoprotein in the red cell membrane from mammalian and avian blood. Carp (Cyprinus carpio) blood was drawn, and prepared the carp red cell membrane. Membrane proteins and glycophorin were examined by SDS- polyacrylamide gel electrophoresis. The Coomassie blue stain pattern of carp red cell membrane was similar to that of rainbow trout and human. The prominent membrane protein, band 3 which designated for human red cell membrane, was detected as well as spectrin, ankyrin and actin. With PAS stain, glycoprotein was detected in the carp red cell membrane. Carp glycophorin was extracted from the membrane by LIS-phenol method, and this preparation showed weak N blood type activity. Sialic acid in carp glycophorin was determined NGNA by thin layer chromatography and other methods. From the result of amino acid analysis, composition was similar to human glycophorin. However, composition of oligosaccharide in carp glycophorin was differing from human oligo
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saccharide. The interaction of carp glycophorin and various lectins showed that the components of carp glycophorin's oligossacharides were O-linked (Gal・GalNAc・Ser/Thr), α Glc and Fuc. After β-elimination, released oligosaccharides from glycophorin were collected, and separated two fractions by using anion exchange column chromatography (HPLC). These fractions were hydrolyzed, following by analysis of high performance anion exchange chromatography with pulsed amperometric detector (HPAEC-PAD). The results from HPAEC-PAD also suggested that the components of oligosaccharides were Gal, Glc and Fuc. Although this results being strengthen by NMR spectra, the structure of these oligosaccharides were not determined without data from GC-MS analysis. We are going to preparation of mg-order desalted oligosaccharides for GC-MS analysis.
From the observation by electron microscope, it seemed that carp glycophorin molecules attached to the flagellum of gram-negative bacterium and to the cell wall of gram-positive bacterium. These results suggested that carp glycophorin has a protective function toward fish pathogen. Less
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