Peterson P A, Rask L, Helting T, Ostberg L, Fernstedt Y
J Biol Chem. 1976 Aug 25;251(16):4986-95.
Crude cell membrane fractions from a number of tissues can form acidic glycolipids. The formation of acidic galactose lipid and mannose lipid was greatly reduced in vitamin A deficiency, primarily in tissues known to be mucus-producing. Mouse mastocytoma tissue was active in forming acidic galactose lipids with UDP-galactose as substrate. One of the products was identified as retinylphosphate galactose. The synthetase reaction producing this compound exhibited an apparent pH optimum at 6.3. The presence of detergent and retinol stimulated the synthetase reaction, which exhibited an absolute requirement for Mn2+ or Mg2+. The synthetase reaction was readily reversible. Incubation of particulate enzyme with retinylphosphate galactose and UDP yielded UDP-galactose and a compound tentatively identified as retinylphosphate. The galactose lipid was isolated by column chromatography on DEAE-cellulose and silica gel. The retinylphosphate galactose was homogeneous when examined by thin layer chromatography. Mild acid hydrolysis of labeled retinylphosphate galactose yields [14C]galactose, whereas alkaline hydrolysis and hydrogenolysis produced [14C]galactose 1-phosphate. Retinylphosphate galactose bound to vitamin A-depleted, retinol-binding protein.
来自多种组织的粗制细胞膜组分能够形成酸性糖脂。在维生素A缺乏的情况下，酸性半乳糖脂和甘露糖脂的形成大幅减少，主要是在已知会产生黏液的组织中。小鼠肥大细胞瘤组织以UDP-半乳糖为底物，在形成酸性半乳糖脂方面具有活性。其中一种产物被鉴定为视黄基磷酸半乳糖。产生这种化合物的合成酶反应在pH 6.3时表现出明显的最佳活性。去污剂和视黄醇的存在刺激了合成酶反应，该反应对Mn2+或Mg2+有绝对需求。合成酶反应很容易逆转。将颗粒酶与视黄基磷酸半乳糖和UDP一起孵育，会产生UDP-半乳糖和一种暂定为视黄基磷酸的化合物。通过在DEAE-纤维素和硅胶上进行柱色谱分离出半乳糖脂。通过薄层色谱法检测时，视黄基磷酸半乳糖是均一的。对标记的视黄基磷酸半乳糖进行温和酸水解会产生[14C]半乳糖，而碱水解和氢解则产生[14C]半乳糖1-磷酸。视黄基磷酸半乳糖与维生素A缺乏的视黄醇结合蛋白结合。
