Exploitation on Biological Activities of Sialoglycosphingolipids and Their Synthetic Family Compounds (Neoglycollipids)

唾液酸鞘糖脂及其合成家族化合物（新糖脂）的生物活性开发

基本信息

批准号：
09359001
负责人：
SAITO Masaki
金额：
$ 20.03万
依托单位：
National Cancer Center Research Institute (1998-1999)Hokkaido University (1997)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

Glycoconjugates from the frontier of cell-to-cell and cell-to-substratum interactions. Among them, gangliosides (sialic acid containing glycosphingolipids) are known to bear important functions in various biological phenomena such as cell growth and differentiation, embryogenesis and carcinogenesis. In vertebrates, almost all the gangliosides are synthesized from a common pre-cursor, ganglioside GM3, which was previously shown by us to exhibit differentiation-inducing activity against human myelogenous leukemia cells. In this project, using the ellaborately-devised expression cloning method, we succeeded for the first time in isolating and molecularly characterizing a new and relevant gene (cDNA and genome) which encodes a key glycosyltransferase, human ganglioside GM3 synthase (sialyltransferase-1: ST3GalV) and then, murine ST3GalV, which is responsible for GM3 biosynthesis. Subsequently, 3 kinds of transcripts (L-,B1-and B2-type) of the gene were detected in mice by 5'-RACE analyses … More whereas a single transcript detectable in human organs, and murine tissue-specific expressions were clarified: L-type transcript was specifically expressed in liver while B1-type was generally detected in various organs with B2-type marginally expressed. The human and murine genomic structures of ST3GalV have been determined by screening BAC and 【lambda bar】 library, respectively, prepared from the chromosomal DNA. Transfection of this enzyme cDNA into ganglioside-deficient mouse lung carcinoma 3LL cells was interestingly shown to introduce the characteristic shedding of GM3-rich membrane domain into the medium. In the programs for exploitation of natural and synthetic sialoglycocompounds, i.e. neoglycolipids, in the applications to the medical fields, the hydrophobic (fatty acid) moiety of ganglioside GM3 was changed in the chemically-synthesized molecule in order to enhance its biological activity, and, among more than 20 synthetic sialoglycolipid compounds, both α-sialo-cholesterol and α-sialodiglyceride were shown to exhibit significant differentiation-including and apoptosis-including activities to human leukemia cells. Anti-sense oligonucleotide therapy for human melanomas is initiated using GD3 synthase cDNA since GD3-dominant melanoma was reported worse in the prognosis. We could produce ST3GalV soluble forms as MBP-/GST-fusion proteins in order to utilize in the development of automatic carbohydrate-chain synthesizers. We have also devised newly the toroidal coil counter-current chromatography to isolate less polar alkali-labile glycolipids and proteins with higher sensitivity. Less

来自细胞到细胞 - 肌间相互作用的糖缀合物。这是我们以前显示出对人髓质白血病细胞的分化诱导活性。人神经节苷脂GM3合成液（siAllltransferase-1：st3galv）和鼠sT3GALV，负责GM3生物合成的鼠。 -race分析...在人体器官和鼠组织特异性表达式中阐明：在肝脏中特异性表达的L型类型类型，在B2型的各种器官中均匀表达。通过筛选BAC和[Lambda Bar]文库确定，从染色体DNA中制备，在培养基中脱离了富含GM3的膜。应用在化学合成的分子中更改了神经节苷脂GM3的医学领域，以增强其生物学活性，并且在20多个合成唾液酸糖脂中，两者都在α-硅氧醇和α-sialo-胆固醇和αα和αα-sialo-sialoglycolipids中。 - 二甘油三酸酯表现出对人类黑色素瘤的雄性白血病（包括人白血病）的尖锐性差异和凋亡。在自动碳水化合物链的发育中，我们还设计了新的曲线盘凝胶电流色谱图，以隔离较小的极性碱性糖脂和蛋白质，并具有较高的敏感性