Role of glycosaminoglycan-synthesizing enzymes in corneal transparency

糖胺聚糖合成酶在角膜透明度中的作用

• 批准号: 14571693
• 负责人: NAKAZAWA Kiyoshi
• 金额: $ 0.96万
• 依托单位: Meijo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571693/
• 关键词: GlcNAc 6-sulfotransferase; Dermatan-specific Gal ; NAc 4-ST Chondroitin synthase; Chondroitin synthase; β-Galactosyltransferase; Chick cornea; cDNA cloning; Glycosaminoglycan; Corneal transparency; GlcNAc sulfotransferase(GlcNAc6ST); Dermatan-spe

cDNAs of some glycosyltransferases and sulfotransferases which are involved in biosynthesis of glycosaminoglycan chain were cloned from chick corneal cells.(2)N-Acetylglucosamine 6-sulfotransferase (GlcNAc6ST) : GlcNAc6ST is involved in a sulfation of GlcNAc residue of keratan sulfate chain. This enzyme could be a marker for the corneal differentiation. Most part (90%) of the sequence of ORF of the cDNA has been determined. The sequence showed 83.0% homology with human GlcNAc6ST-1, but not with human cornea-specific GlcNAc6ST-5. Furthermore, 300 bps of 5'-region of the sequence showed very low homology even with the GlcNAc6ST-1.(3)Chondroitin synthasae and β-glucosyltransferases (Gal-T) : Chondroitin synthase is involved in the elongation of chondroitin chain and Gal-T in the elongation of keratan sulfate chain. Both enzymes could be marker for the dedifferentiation or transdifferentiation from cornea. The sequences of ORFs of both cDNAs have been determined. The sequence of chick chondroitin synthase cDNA showed 82.0% homology with human chondroitin synthase-1, while that of chick Gal-T was identical with chick Gal-T I reported previously. Expression levels of these two enzymes in cultured stromal cells were examined by real-time PCR method. The levels of the two enzymes did not changed so much during the passage of culture (tertiary culture).

从鸡角膜细胞中克隆了与糖胺聚糖链生物合成有关的糖基转移酶和磺基转移酶的cDNA。（2）N-乙酰葡糖胺6-磺基转移酶（GlcNAc 6ST）：GlcNAc 6ST参与硫酸角质素链的GlcNAc残基的硫酸化。该酶可作为角膜分化的标志物。该cDNA的大部分（90%）ORF序列已被确定。该序列与人GlcNAc 6ST-1的同源性为83.0%，但与人角膜特异性GlcNAc 6ST-5的同源性不高。此外，该序列的5 ′-区的300 bp甚至与GlcNAc 6ST-1显示出非常低的同源性。（3）软骨素合成酶和β-葡萄糖基转移酶（Gal-T）：软骨素合成酶参与软骨素链的延长，Gal-T参与硫酸角质素链的延长。这两种酶可作为角膜脱分化或转分化的标志物。已测定了这两个cDNA的ORF序列。鸡软骨素合成酶cDNA序列与人软骨素合成酶-1的同源性为82.0%，鸡Gal-T与已报道的鸡Gal-T Ⅰ的同源性为82.0%。用实时荧光定量PCR方法检测两种酶在培养的基质细胞中的表达水平。这两种酶的水平在传代培养（三级培养）过程中变化不大。

项目成果

Nana Ogata, Ikuko Takahashi, Kiyoshi Nakazawa: "Purification Characterization of Chick Corneal β-D-Glucuronyltransferase Involved in Chondroitin Sulfate Biosynthesis."Biol.Pharm.Bull.. 25(10). 1282-1288 (2002)

Nana Ogata、Ikuko Takahashi、Kiyoshi Nakazawa：“硫酸软骨素生物合成中涉及的鸡角膜 β-D-葡萄糖醛酸基转移酶的纯化特征。”Biol.Pharm.Bull.. 25(10) (2002)。

Susumu Sashima: "Identification and Characterization of β-D-Galactosyltransferase in Chick Cormeas"Jpn.J.Ophthalmol.. 46. 607-615 (2002)

Susumu Sashima：“小鸡中 β-D-半乳糖基转移酶的鉴定和表征”Jpn.J.O善眼科.. 46. 607-615 (2002)

Susumu Sashima, Ikuko Takahashi, Kiyoshi Kanazawa: "Identification and Characterization of b-D-Galactosyltransferase in Chick Corneas"Jpn.J.Ophthalmol.. 46. 607-615 (2002)

Susumu Sashima、Ikuko Takahashi、Kiyoshi Kanazawa：“鸡角膜中 b-D-半乳糖基转移酶的鉴定和表征”Jpn.J.O善眼科.. 46. 607-615 (2002)

Nana Ogata, Ikuko Takahashi, Kiyoshi nakazawa: "Purification and Characterization of Chick Corneal β-D-Glucuronyl-transferase Involved in Chdoritin Sulfate Biosynthesis."Biol.Pharm.Bull. 25(10). 1282-1288 (2002)

Nana Ogata、Ikuko Takahashi、Kiyoshi nakazawa：“参与硫酸 Chdoritin 生物合成的鸡角膜 β-D-葡萄糖醛酸基转移酶的纯化和表征。”Biol.Pharm.Bull 25(10)。

Susumu Sashima, Ikuko Takahashi, Kiyoshi Nakazawa: "Identification Characterization of β-D-Galactosyltransferase in Chick Corneas."Jpn.J.Ophthalmol.. 46. 607-615 (2002)

Susumu Sashima、Ikuko Takahashi、Kiyoshi Nakazawa：“小鸡角膜中 β-D-半乳糖基转移酶的鉴定特征。”Jpn.J.Oleadingmol.. 46. 607-615 (2002)