| Project/Area Number |
11671762
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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 |
Ophthalmology
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| Research Institution | Meijo University |
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Principal Investigator |
NAKAZAWA Kiyoshi Meijo University, Faculty of Pharmacy, Associate Professor, 薬学部, 助教授 (60076750)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Ikuko Meijo University, Faculty of Pharmacy, Technician, 薬学部, 教務技師補 (10200784)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2001: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | N-Acetyglucosaminyl sulfotransferase / β-Glucuronyltransferase / β-Galactosyltransferase / Keratan sulfate / Chondroitin sulfate / Chondroitin 4-sulfotransferase / Chondroitin 6-sulfotransferase / Chick cornea / chondroitin sulfate / chondroitin 4-sulfotransferase / chondroitin 6-sulfotransferase / chick cornea / cDNA cloning / N-acetylglucosamine 6-sulfotransfersase / chondroitin synthase / keratan sulfate / N-Acetylglucosaminyl sulfotransferase / GlcNAc-6-O-sulfotransferase / corneal transparency / corneal proteoglycans / glycosyltransferases / corneal diseases |
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| Research Abstract |
1. Immunohistochemically, keratan sulfate (KS) was first detected in corneal stroma on Day 6 of chick embryonic age, but this KS was undersulfated. The sulfation of KS increased on Day 8.
2. When the stromacytes were cultured in vitro, the degree of sulfation of KS decreased markedly but the syntheses of KS backbone and core protein of KS proteoglycan were maintained. On the other hand, the sulfation of chondroitin sulfate increased.
3. N-Acetylglucosaminyl sulfotransferase which may be involved in the sulfation of KS, was purified 887-fold with a 8.4% yield from 2-day-old chick corneas. The enzyme specifically transferred a sulfate to GlcNAc at the nonreducing end but not to internal GlcNAc. When the stromacytes were cultured in vitro, the activity of this enzyme specifically, markedly decreased.
4. β-Glucuronyltransferase which may be involved in the elongation of chondroitin chain, was purified 389-fold with a 11% yield from 2-day-old chick corneas. This enzyme activity toward chondro-heptasaccharide was 10-fold higher than that toward chondro-pentasaccharide.
5. β-Galactosyltransferase activity toward 6-sulfo-GlcNAcβl-3Gal which is the disaceharide unit of KS, was found in chick corneas. This enzyme may be a novel β-galactosyltransferase.
6. Almost a whole of open reading frame (ORF) of cDNA of chick corneal chondroitin 6-sulfotransferase was determined for the nucleotide sequence and the predicted amino acid sequence, 3'-Side half of ORF of cDNA of chick coraeal chondroitin 4-sulfotransferase was determined for the nucleotide sequence and the predicted amino acid sequence.
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