On the occurrence of diphosphoinositol in the lipids of liver and pancreas

George A. Kfoury, Stanley E. Kerr
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引用次数: 7

Abstract

  • 1.

    1. The lipids of liver were reexamined for polyphosphoinositide, previously reported absent. The presence of a diphosphoinositide (DPI) was revealed by the isolation of inositol diphosphate (IP2) from an acid hydrolysate, and its glyceryl derivative from an alkaline hydrolysate of the concentrated phospholipid fraction.

  • 2.

    2. Starting with lipids extracted from liver and pancreas by azeotropic dehydration with 1,2-dichloroethane, materials soluble in acetone and alcohol were removed. After partition of the alcohol-insoluble phospholipids in CCl4-light petroleum-methanol-H2O (31:31:35:3, V/V), the non-polar fraction was assayed for inositol polyphosphate by chromatographic fractionation of the acid hydrolysate on Dowex-2 X8 Cl A diphosphate mixture eluted by NaCl or LiCl was separated by refractionation with a formate system yielding IP2 and glycerol diphosphate.

  • 3.

    3. The mixed glyceryl derivatives of IP2 and glycerol diphosphate were secured by chromatographic fractionation of the alkaline hydrolysate. Based on the recovery of IP2 and its glyceryl derivative, the content of DPI in the original pork liver lipid is 0.28 mmoles/kg.

  • 4.

    4. DPI could not be extracted by ether from acetone-dehydrated fresh beef liver but was extracted with neutral chloroform-methanol (2:1, V/V).

  • 5.

    5. Most of the IP2 found in pacreatic lipid occured in material insoluble in CHCl3 and in the CCl4-light petroleum solvent.

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肝脏和胰腺脂质中二磷酸肌醇的出现
1.1. 重新检查肝脏脂质是否有先前报道不存在的多磷酸肌醇。二磷酸肌醇(IP2)从酸性水解物中分离得到,其甘油衍生物从浓缩磷脂部分的碱性水解物中分离得到,结果表明二磷酸肌醇(DPI)存在。从用1,2-二氯乙烷共沸脱水法提取肝脏和胰腺脂质开始,去除可溶于丙酮和醇的物质。在ccl4 -轻质石油-甲醇- h2o (31:31:35:3, V/V)中对醇不溶性磷脂进行分离后,将酸水解产物在Dowex-2 X8 Cl上进行色谱分馏,测定非极性部分的肌醇多磷酸;用NaCl或LiCl洗脱的二磷酸混合物,在生成IP2和二磷酸甘油的甲酸体系中进行折射分离。通过对碱性水解产物的色谱分离,获得了IP2和二磷酸甘油的混合甘油衍生物。通过对IP2及其甘油衍生物的回收,得到原猪肝脂中DPI的含量为0.28 mmol /kg.4.4。丙酮-脱水鲜牛肝不能用乙醚法提取DPI,可用中性氯仿-甲醇(2:1,V/V)萃取。脂质中发现的大部分IP2发生在不溶于CHCl3的物质和ccl4 -轻石油溶剂中。
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