2000 Fiscal Year Final Research Report Summary
Molecular Cloning of E-Selectin-Glycoprotein Counter Ligand, gp150.
| Project/Area Number |
11670147
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pathological medical chemistry
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| Research Institution | JICHI MEDICAL SCHOOL |
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Principal Investigator |
NAKAMURA Mitsuru Jichi Med.Sch., Med., Lecturer, 医学部, 講師 (20198237)
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| Project Period (FY) |
1999 – 2000
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| Keywords | E-Selectin / Sialyl-LeX / Counter Ligand / Cell Adhesion / Sialomucin Family / O-Linked Glycoprotein |
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| Research Abstract |
In human pre-B leukemia cells sialyl-LeX (sLeX) antigen is not expressed on every protein but expressed on a specific core glycoprotein with molecular size of 150kDa (gp150). In addition, cell surface sLeX antigen expression level is controlled by core 2 N-acetylglucosaminyltransferase (C2GnT) gene. In addition, we have revealed the followings.
1. Not only in pre-B cell differentiation but also in tonsillar B cell activation, C2GnT regulates cell surface sLeX expression level.
2. Anti-sLeX mAb reactivity of gp150 was disappeared after O-sialoglycoprotein-endopeptidase (from Pasteurella haemolytica) treatment. This suggests gp150 must be a sialomucin type glycoprotein molecule that has abundant O-linked sugar chains with sialic acid on their termini.
3. A tryptic peptide was purified and analyzed from pre-B NALL-1 cell line membranous fraction using anti-sLeX mAb-conjugated protein-G.The peptide was determined as a mucin-type glycoprotein. It was confirmed by immunostaining methods.
4. The cDNA of gp150 was obtained and transfected in the antisense orientation into a sLeX-highly expressed pre-B cell line. Sialyl-LeX-reactivity in the transfected cell line was significantly downregulated and E-selectin-dependent cell adhesion capability was remarkably reduced.
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[Publications] Nakamura M, Furukawa Y, Sasaki R, Masuyama J, Kikuchi J, Iwase S, Kudo T, Narimatsu H, Asakura S, Fujiwara S, and Inokuchi J.: "UDP-GlcNAc : Galβ1→3GalNAc (GlcNAc to GalNAc) β1→6N-acetylglucosaminyltransferase Holds a Key Role on the Control of CD15s Expression in Human pre-B Lymphoid Cell Lines."Glycobiology. 9. 1-12 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Yang, W., Asakura, S., Sakai, T., Nakamura, M., Fujimura, K., and Matsuda, M.: "Two-step Spreading Mode of Human Glioma Cells on Fibrin Monomer via a β1 Integrin : Interaction with Its Ligand in the Substratum and with Endogenous Cellular Fibronectin in the Extracellular Matrix."Thromb.Res.. 93. 279-290 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Furukawa, Y., Iwase, S., Kikuchi, J., Nakamura, M., Yamada, H., and Matsuda, M.: "Transcriptional Repression of the E2F-1 Gene by Interferon-a Is Mediated through Induction of E2F-4/pRB and E2F-4/p130 Complexes."Oncogene. 18. 2003-2014 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Iwase, S., Furukawa, Y., Kikuchi, J., Saito, S., Nakamura, M., Nakayama, R., Horiguchi-Yamada, J., and Yamada, H.: "Defective binding of IRFs to the initiator element of interleukin-1β-converting enzyme (ICE) promoter in an interferon-resistant Daudi subline."FEBS Lett. 450. 263-267 (1999)
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[Publications] Muroi K, Tarumoto T, Akioka T, Kirito K, Nagai T, Izumi T, Nakamura M, Hatake K, Hakomori S, Miura Y, and Ozawa K.: "Sialyl-Tn- and Neuron Specific Enolase-Positive Minimally Differentiated Erythroleukemia."Internal Medicine. 39(10). 843-846 (2000)
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