Modifications of Alkaline Phosphatase Molecules by the Glycan PI-Anchor and Their Applications for Pathological Discrimination

聚糖 PI-Anchor 对碱性磷酸酶分子的修饰及其在病理判别中的应用

• 批准号: 02671072
• 负责人: KOMODA Tsugikazu
• 金额: $ 1.34万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02671072/
• 关键词: Alkaline Phosphatase; GPI-anchor; PIPLC; Diabetic rats; Intestinal enzyme; Stearate; Modification; Susceptibility; ペプチド-アンカ-; Kasaharaアイソザイム; 癌化

It has been revealed that the C-terrminal amino acids of Alkaline phosphatase (ALP) molecules, except for the intestinal ALP, link to glycanphosphoinositol (GPI) involving ethanolamine and 1, 2-diacylglycerol of the GPI inserts to the biomembrane. In this regard, human placental ALP is released from the plasma membrane by phosphatidylinositol-specific phospholipase C (PIPLC), while the rat intestinal ALP is not released by this treatment (seetharam, B. ET AL. : arch. biochem. biophys., 253 : 189-l98, 1987). The fact suggests that the manner in which the GPI anchors with these ALPS is different.With the streptozotocin-induced diabetic rats, the major ALP detected in serum was intestinal ALP. In vitro, ALP was readily released from the duodenum by piplc, but little if any was released from the ileum. The characteristics of serum ALP in rats is similar to the ileal ALP, suggesting that the circulating ALP is derived from the ileum. As reported with the IGG receptor, therefore the intestinal ALP from rats exist in both peptide-anchored and GPI-anchored forms. Other possibility, the 11eal enzyme may bears the acylated form, as human cholineesterase.On the other hand, rabbit kidney have predominantly intestinal ALP, unlike humans. The intestinal ALP in rabbit kidney was clearly released by PIPLC treatment, like the duodenal ALP in rats. Chemical Analyses revealed that the purified intestinal ALP contains palmitate, stearate, and inositol. Molar ratios of three components were calculated to be 1 MOL, respectively. The stearate content of intestinal ALP from methylnitrosourea-induced cancer tissues of rats was calculated to be 2 MOL, the intestinal ALP in the cancer tissues was resistant to PIPLC treatment, indicating that modification of the GPI moiety in intestinal ALP molecules may alter the susceptibility to PIPLC.

据透露，除肠道ALP外，碱性磷酸酶（ALP）分子的C-杂氨酸氨基酸与涉及乙醇胺的糖磷酸肌醇（GPI）的联系，涉及乙醇胺和1、2-二酰基甘油的GPI插入到生物膜的GPI插入。在这方面，磷脂酰肌醇特异性磷脂酶C（PIPLC）从质膜中释放人胎盘ALP，而这种治疗方法尚未释放大鼠肠道ALP（Seetharam，B。等人：Arch。B.等：Arch。Biophem。Biophys。Biophys。 253：189-L98，1987）。事实表明，用这些ALP锚定GPI的方式不同。链蛋白酶诱导的糖尿病大鼠，血清中检测到的主要ALP是肠道ALP。在体外，ALP很容易被PIPLC从十二指肠中释放出来，但几乎没有释放出回肠。大鼠血清ALP的特征与回肠ALP相似，这表明循环ALP源自回肠。正如IgG受体报道的那样，因此来自大鼠的肠道ALP都存在于肽锚定和GPI锚定的形式中。另一种可能性，这种酶可能具有酰化形式，因为人类胆固醇酶。另一方面，兔肾脏主要具有肠道ALP，与人不同。 PIPLC治疗清楚地释放了兔肾脏中的肠道ALP，例如大鼠的十二指肠ALP。化学分析表明，纯化的肠道ALP含有棕榈酸酯，硬化酸盐和肌醇。计算出三个组分的摩尔比分别为1 mol。从甲硝基甲基甲基催化的大鼠癌组织中的肠道ALP的硬脂酸含量计算为2摩尔，癌症组织中的肠道ALP对PIPPC治疗具有抗性，表明在肠道ALP分子中修饰GPI部分可能会改变易感性以改变易感性PIPLC。

项目成果

A.Nagata,N.Hirota,T.Sakai,M.Fujimoto,T.Komoda: "Molecular nature and possible presence of a membranous glycan-phos-phatidylinositol anchor of CA 125 antigen." Tumor Biology. 12. 279-286 (1991)

A.Nagata、N.Hirota、T.Sakai、M.Fujimoto、T.Komoda：“CA 125 抗原的膜聚糖-磷脂酰肌醇锚的分子性质和可能存在。”

菰田 二一,三浦 雅一,小山 岩雄(分担): "電気泳動法のすべて(Medical Technology別冊)" 医歯薬出版株式会社(桜林郁之介,熊坂一成編), 9 (1991)

小茂田新一、三浦正一、小山岩生（撰稿人）：《关于电泳的一切（医疗技术特刊）》石药出版株式会社（樱林育之介、熊坂一成编），9（1991）

S.Unakami,T.Komoda,Y.Sakagishi: "Translocation of intestinal alkaline phosphatase in streptozotocininduced diabetic rats." International Journal of Biochemistry. 22. 1325-1331 (1990)

S.Unakami、T.Komoda、Y.Sakagishi：“链脲佐菌素诱导的糖尿病大鼠肠道碱性磷酸酶的易位。”

坂井 隆夫,牧田 明,近藤 朝明,滝川 一,三宅 和彦,山中 正己,菰田 二一,坂岸 良克: "Phosphatidylinositolーspecific phospholipase C処理した血清アルカリ性ホスファタ-ゼアイソザイムのpolyacrylamideーgel disc電気泳動法による検討" 医学のあゆみ. 155. 741-742 (1990)

Takao Sakai、Akira Makita、Tomoaki Kondo、Hajime Takikawa、Kazuhiko Miyake、Masami Yamanaka、Niichi Komoda、Yoshikatsu Sakagishi：“用磷脂酰肌醇特异性磷脂酶 C 处理血清碱性磷酸酶同工酶的聚丙烯酰胺凝胶盘电泳”“历史回顾医学 155. 741-742 (1990)

I.Koyama,Y.Fujimori Arai,N.Hirota,T.Sakai,Y.Sakagishi,T.Komoda: "Liver-like alkaline phosphatase in the tissue-unspecific type enzyme found in rabbit organs." Biochimica et Biphysica Acta. 1080. 165-172 (1991)

I.Koyama、Y.Fujimori Arai、N.Hirota、T.Sakai、Y.Sakagishi、T.Komoda：“兔器官中发现的组织非特异性酶中的肝脏样碱性磷酸酶。”