Comprehensive studies on N-and O-glycans expressed on different stage of cells
| Project/Area Number |
17390012
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | Kinki University |
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Principal Investigator |
KAKEHI Kazuaki Kinki University, Faculty of pharmaceutical Sciences, Professor (30101405)
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| Co-Investigator(Kenkyū-buntansha) |
KINOSHITA Mitsuhiro Kinki University, Faculty of pharmaceutical Sciences, Assistant Professor (40330279)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥9,660,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥660,000)
Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2005: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | Glycome / N-linked glycan / Cancer cell / Tumor marker / Polylactosamine-type glycan / Free glycan / Stomach cancer cell / N結合型糖鎖 / グライコフォーム / 胃癌 |
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| Research Abstract |
In this research project, we developed a method for comprehensive analysis of N- and 0-linked glycans present on cell membrane. The method was applied to comparative analysis of total glycans expressed on various cancer cell surfaces. The total N-glycan pool derived from cell membrane was fractionated into five fractions based on the number of sialic acids by serotonin-affinity chromatography. After digestion with sialidase, each fraction was further separated by normal-phase HPLC, and structures of oligosaccharide were analyzed by the combination of MALDI-TOF MS with exoglycosidase digestion. High mannose-type and complex-type glycans were commonly observed in all cancer cells examined in the present study. In contrast, characteristic oligosaccharides were also observed in some cancer cells. We found that U937 cells contained a large amount of polylatosamine-type N-glycans and MKN45 cells expressed perfucosylated N-glycans. Presence of these characteristic glycan strongly suggests the
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expression of the related enzyme involved in their biosynthesis.
During the studies on the comparative analysis of total N-linked glycans of cellular membrane, we found that characteristic free glycans were accumulated in MKN45 and MKN7 cells. All free glycans observed in both cells contained only one GlcNAc residues at the reducing end and the most abundant free glycan found in these cells was determined to be NeuAcα2-6Ga1β1-4G1cNAcβ1-2Manαl-3Manβ1-4G1cNAc Biochemical analyses indicated that the characteristic oligosaccharide was cytosolic glycans derived from lysosomes due to the low integrity of lysosomal membrane. Since the accumulation of such free N-glycans was specific to these cells among various cancer cell lines examined, these cytosolic free glycans may serve as a specific biomarker for diagnosis of specific tumors.
The technique developed in this project will be a powerful tool for comparative glycomics studies in cells, and has a big potential for contributing to exploration of a new post-proteomics era, glycomics, and eventually for breakthrough in healthcare including establishment of diagnosis and therapies for various diseases. Less
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Report
(4 results)
Research Products
(133 results)
