Analysis of membrane・bound form in alkaline phosphatase induced by short chain fatty acid.

短链脂肪酸诱导的碱性磷酸酶的膜结合形式分析。

• 批准号: 07672403
• 负责人: NAKABAYASHI Toshikatsu
• 金额: $ 1.22万
• 依托单位: Mukogawa Women's University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672403/
• 关键词: short chain fatty acied; alkaline phosphatase; membrane-bound form

The results of this project are as follows :1. When cultured cells or tissues which express kidney, bone or small intestinal types of alkaline phosphatase (ALP) were treated with phosphatidylinositol・specific phospholipase C (PI-PLC) or glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD), ALP was completely solubilized.2. Among the HeLa cells tested, HeLa 229 cells showed the highest ability of ALP induction by sodium butyrate (SB). This ALP induction is considered from a change in the enzymatic property. Using a specific antibody and cell sorter, the present site of newly synthesized ALP was whown to be the cell surface. Furthermore, ALP activity was suppressed in the presence of reagents which inhibit the transcription or protein synthesis. These data indicate that ALP induction in HeLa 229 cells by SB is due to changes in the transcriptional level and amount in the synthesis.3. When HeLa 229 cells were treated with PI-PLC or GP-PLD,the ratio of solubilization of ALP with or without SB treatment was about 10-30 or 50%, respectively. These findings indicate that ALP forms having different sensitivities against PI-PLC or GPI-PLD are present in HeLa 229 cells.These findings indicate that the membrane-bound form of ALP in HeLa 229 cells expressed by placental type shows diversity compared with those of cultured cells or tissues expressed by kidney, bone or small intestinal types of ALP.

本课题的主要研究成果如下：1.用磷脂酰肌醇·特异性磷脂酶C（PI-PLC）或糖基化磷脂酰肌醇特异性磷脂酶D（GPI-PLD）处理表达肾型、骨型和小肠型碱性磷酸酶（ALP）的培养细胞或组织，ALP被完全溶解.在所测试的HeLa细胞中，HeLa 229细胞显示出丁酸钠（SB）诱导ALP的最高能力。这种ALP诱导被认为是酶性质的变化。利用特异性抗体和细胞分选仪，确定新合成的ALP的存在部位为细胞表面。此外，ALP活性在抑制转录或蛋白质合成的试剂的存在下被抑制。这些数据表明SB对HeLa 229细胞ALP的诱导是由于转录水平和合成量的变化.当用PI-PLC或GP-PLD处理HeLa 229细胞时，SB处理或不处理的ALP的增溶率分别为约10-30或50%。这些发现表明，在HeLa 229细胞中存在对PI-PLC或GPI-PLD具有不同敏感性的ALP形式，这些发现表明，与表达肾、骨或小肠型ALP的培养细胞或组织相比，表达胎盘型ALP的HeLa 229细胞中的ALP的膜结合形式显示出多样性。

项目成果

Chisako Itami: "Growth inhibition, morphological change and ectoenzyme release of LLC-PK1 cells by phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis." Biosci, Biotech.Biochem.(in press).

Chisako Itami：“苏云金芽孢杆菌磷脂酰肌醇特异性磷脂酶 C 对 LLC-PK1 细胞的生长抑制、形态变化和胞外酶释放。”

Chisako Itami: "Release of ectoenzymes from small intestine brush border membranes of mice by phospholipases." Biosci.Biotech.Biochem.61(2). 336-340 (1997)

Chisako Itami：“磷脂酶从小鼠小肠刷状缘膜中释放胞外酶。”

Chisako Itami: "Growrh inhibition morphological change and ectoenzyme release of LLC-PK1 cells by phosphatiodyinositol-specific phopholiolipase C of Bacilus thuringiensis." Biosci.Bitoech.Biochem.(in press). (1997)

Chisako Itami：“苏云金芽孢杆菌的磷酸肌醇特异性磷脂酶 C 对 LLC-PK1 细胞的生长抑制、形态变化和胞外酶释放。”

Chisako Itami: "Release of ectoenzymes from small intestine brush border membranes of mice by phospholipases." Biosci.Biotech.Biochem.61・2. 336-340 (1997)

Chisako Itami：“磷脂酶从小鼠小肠刷状缘膜中释放胞外酶。”Biosci.Biotech.Biochem.61·2（1997）。

Chisako Itami: "Release of ectoenzymes from small intestine brush border membranes of mice by phospholioases." Biosci.Bitoech.Biochem.61・2. 336-340 (1997)

Chisako Itami：“通过磷酸酶从小鼠小肠刷状缘膜中释放胞外酶。”Biosci.Bitoech.Biochem.61·2（1997）。