2002 Fiscal Year Final Research Report Summary
Biological significance of N-acetylglucosamine 6-sulfation
| Project/Area Number |
12480187
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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 |
Functional biochemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
MURAMATSU Takashi Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (00030891)
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| Co-Investigator(Kenkyū-buntansha) |
MURAMATSU Hisako Nagoya University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 講師 (50182134)
KADOMATSU Kenji Nagoya University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (80204519)
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| Project Period (FY) |
2000 – 2002
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| Keywords | sulfotransferase / N-acetylglucosamine / L-selectin / Blood coagulation / Oligosaccharide synchesis / Glycoproteins |
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| Research Abstract |
Three species of N-acetylglucosamine-6-sulfotransferases (GlcNAc6ST), namely GlcNAc6ST-1, GlcNAc6ST-2, GlcNAc6ST-3 were expressed and the specificities were compared. GlcNAc6ST-l as well as GlcNAc6ST-2 acted on mucin-type core 2 sugar (GlcNAcβ1-6[Galβ1-3]GalNAc) and there were not significant differences between the two enzymes. Although GlcNAc6ST-2 is known to be involved in lymphocyte homing, it is not clarified whether GlcNAc6ST-1 plays such a role. Cells doubly transfected with GlcNAc6ST-1 and fucosyltransferase VII induced roling of L-selectin-expressing cells. GlcNAc6ST-3 acted only on core 2 sugars. On the other hand, only GlcNAc6ST-2 acted on core 3 sugars (GlcNAcβ1-3 GalNAc). From this specificity, GlcNAc6ST-2 was considered to be the enzyme, which became newly expressed in mucinous adenocarcinoma, and this point was confined by RT-PCR analysis.
We produced knockout mice deficient in the GlcNAc6ST-1 gene. That this mice completely lacked GlcNAc6ST-1 was confirmed by Southern and Northern blot analyses. The mice was born normally, and showed no apparent abnormality. Currently backcross is on the way, so that we are able to analyze possible abnormality in lymphocyte homing.
GilcNAc6ST-1 was produced as fasion protein in Tn5 cells using the Baculovirus system. The purified recombinant enzyme was useful for synthesis of sulfated oligosaccharides and sulfation of glycoprotein-bound carbohydrates.
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