Joint Research on Acidic Glycoconjugates on Cell Surfaces

细胞表面酸性糖复合物的联合研究

• 批准号: 10044265
• 负责人: KITAJIMA Kenji
• 金额: $ 4.22万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10044265/
• 关键词: POLYSIALIC ACID; OLIGOSIALIC ACID; KDN; SULFATED GLYCAN; FERTILIZATION; SURFACE GLYCANS TIMOR CELLS; METABOLISM OF SIALIC ACID; GLYCOLIPID-ENRICHED MICRODOMAIN; 神経細胞接着分子

The objective of this research project was to elucidation of regulatory mechanism for cell recognition and adhesion by acidic glycan chains of cell surface glycoconjugates. The results follow :1. Clarification of biosynthetic mechanism of oligo/polysialic acid units in brain and nerve system : (1) Several glycoproteins were shown to contain oligo/polysialic acid structures in fetal and adult mammalian brains. Studies on role of this epitope in biological activities in nerve cell system are now underway ; (2) More than 10 new monoclonal antibodies that differentially recognized various di/oligosialic acid with different chain lengths were prepared as probes for function/structure studies. Two highly sensitive chemical methods to detect di/oligosialic acid using fluorescent dye were developed.2. Role of KDN-glycans or sulfated sialoglycans in gamete interactions at fertilization : (1) The low density detergent-insoluble membrane fractions which were rich in glycolipids were prepared and characterized for unique features of lipid and protein compositions. This membrane fractions are suggested to mediate sperm-egg interaction as adhesion units on the cell surface ; (2) Three distinct cDNAs for sialyltransferases that are involved in synthesis of polysialic and in fish oocyte were cloned ; (3) An avian egg envelope acidic glycoprotein (ZPC) was shown to be biosynthesized in granulosa cells of follicles.3. Expression and roles of polySia- or KDN-containing glycan chains in tumor cells : (1) KDN-glycan chains were shown to be a oncodevelopmental antigen ; (2) Elevated expression of intracellular KDN in mammalian cells cultured in mannose-rich medium was observed. Some cancer cells whose KDN level is high may have alternations of mannose metabolism ; (3) A cDNA for CMP-KDN synthetase from trout testis was successfully cloned. Substrate specificity with KDN of this type of enzyme is suggested to determine the level of KDN in mammalian normal and malignant cells.

该研究项目的目的是阐明细胞表面糖复合物的酸性聚糖链对细胞识别和粘附的调节机制。结果如下： 1．大脑和神经系统中寡聚/聚唾液酸单元的生物合成机制的阐明：(1)在胎儿和成年哺乳动物大脑中，几种糖蛋白被证明含有寡聚/聚唾液酸结构。目前正在研究该表位在神经细胞系统生物活性中的作用； (2)制备了10余种差异识别不同链长的二/低聚唾液酸的新型单克隆抗体作为功能/结构研究的探针。开发了两种利用荧光染料检测二/低聚唾液酸的高灵敏度化学方法。 2． KDN-聚糖或硫酸化唾液酸聚糖在受精时配子相互作用中的作用：(1)制备富含糖脂的低密度去污剂不溶性膜级分，并表征脂质和蛋白质组合物的独特特征。该膜部分被认为作为细胞表面上的粘附单位来介导精卵相互作用； (2) 克隆了参与多唾液酸合成和鱼卵母细胞中唾液酸转移酶的三种不同的cDNA； (3)卵泡颗粒细胞中可生物合成禽卵包膜酸性糖蛋白(ZPC)。3．含polySia或KDN的聚糖链在肿瘤细胞中的表达和作用：（1）KDN-聚糖链被证明是一种肿瘤发育抗原； (2)在富含甘露糖的培养基中培养的哺乳动物细胞中观察到胞内KDN表达升高。一些KDN水平较高的癌细胞可能会出现甘露糖代谢的改变； (3)成功克隆了鳟鱼睾丸CMP-KDN合成酶的cDNA。建议利用此类酶的 KDN 的底物特异性来确定哺乳动物正常和恶性细胞中 KDN 的水平。

项目成果

Takashi Angata: "Biosynthesis of KDN. Identification and characterization of a KDN-9-phosphate synthetase activity from trout testis"J. Biol. Chem.. 274. 22949-22956 (1999)

Takashi Angata：“KDN 的生物合成。鳟鱼睾丸中 KDN-9-磷酸合成酶活性的鉴定和表征”J。

Takashi Angata: "Elevated expression of free deaminoneuraminic acid in mammalian cells cultured in mannose-rich media"Biochem. Biochem. Res. Commun.. 261. 326-331 (1999)

Takashi Angata：“在富含甘露糖的培养基中培养的哺乳动物细胞中游离脱氨基神经氨酸的表达升高”Biochem。

Angata, Takashi: "Biosynthesis of KDN. Identification and characterization of a KDN-9-phosphate synthetase activity from trout testis"J. Biol. Chem.. 274. 22949-22956 (1999)

Angata，Takashi：“KDN 的生物合成。鳟鱼睾丸中 KDN-9-磷酸合成酶活性的鉴定和表征”J。

Kitajima, Ken: "Occurrence of α2→8-linked oligosialic acid residues in mammalian glycoproteins"Sialobiology and Other novel Form of Glycosylation. 69-76 (1999)

Kitajima，Ken：“哺乳动物糖蛋白中α2→8-连接的寡唾液酸残基的出现”Sialobiology and Other new Form of Glycosylation 69-76 (1999)。

Ohta, Kaoru: "Isolation and characterization of low density detergent-insoluble membrane (LD-DIM) fraction from sea urchin sperm"Biochem. Biophys. Res. Commun.. 258. 616-623 (1999)

Ohta，Kaoru：“海胆精子中低密度去污剂不溶性膜 (LD-DIM) 组分的分离和表征”Biochem。