Chana R S, Treleaven W D, Cushley R J, Steinbrecher U P
Department of Chemistry, Simon Fraser University, Burnaby, B.C., Canada.
Biochem Cell Biol. 1990 Jan;68(1):180-8. doi: 10.1139/o90-024.
Selectively labelled lipids have been incorporated into the surface monolayer of human serum low density lipoprotein (LDL) and very low density lipoprotein (VLDL). From 3 to 17 mol% of phosphatidylcholine, selectively deuterated at various positions along the sn-2-acyl chain, was transferred from unilamellar vesicles to VLDL using a partially purified phosphatidylcholine transfer protein. Selectively deuterated palmitic acids were incorporated into LDL (6-20 mol%) and into VLDL (7-10 mol%). Electron microscopy, light scattering, and 31P nuclear magnetic resonance indicated that particle size remained unchanged. Gel exclusion chromatography and chemical analysis showed no difference in hydrodynamic properties and only slight alteration to particle component ratios. Biological activity of labelled VLDL was measured from the rate of cholesterol esterification by cultured J774A.1 cells. Effect of labelling LDL was evaluated by monitoring LDL uptake and degradation by cultured human skin fibroblasts. In all cases the lipoproteins containing labels were indistinguishable from their native counterparts.
已将选择性标记的脂质掺入人血清低密度脂蛋白(LDL)和极低密度脂蛋白(VLDL)的表面单层中。使用部分纯化的磷脂酰胆碱转移蛋白,将沿着sn-2-酰基链在不同位置选择性氘代的3至17摩尔%的磷脂酰胆碱从单层囊泡转移至VLDL。将选择性氘代的棕榈酸掺入LDL(6-20摩尔%)和VLDL(7-10摩尔%)中。电子显微镜、光散射和31P核磁共振表明颗粒大小保持不变。凝胶排阻色谱法和化学分析表明,流体动力学性质没有差异,颗粒组分比例仅有轻微改变。通过培养的J774A.1细胞的胆固醇酯化速率来测量标记的VLDL的生物活性。通过监测培养的人皮肤成纤维细胞对LDL的摄取和降解来评估标记LDL的效果。在所有情况下,含有标记的脂蛋白与其天然对应物没有区别。