Trepanier D J, Thibert R J
Department of Chemistry and Biochemistry, University of Windsor, Ontario, Canada.
Clin Biochem. 1996 Aug;29(4):333-45. doi: 10.1016/0009-9120(96)00018-5.
To study the binding of cyanate to erythrocyte membrane aminophospholipids in vitro, and to investigate whether carbamylated aminophospholipids can be detected in the plasma membrane of native erythrocytes.
For in vitro studies, the lipid components of 14C-carbamylated erythrocyte membranes were resolved by thin-layer chromatography (TLC). The covalent incorporation of cyanate was visualized by autoradiography and quantitated by phosphorus analysis. For the in vivo studies, phospholipid headgroups were enzymatically hydrolyzed by phospholipase D and subsequently reacted with diacetyl monoxime.
Both phosphatidylethanolamine (PE) and phosphatidylserine (PS) were covalently modified by [14C] cyanate; incorporating 15.76 +/- 0.09 and 13.34 +/- 0.81 mol%, respectively, following a 15-h incubation. Carbamylated PE (carb-PE) was resolved with PE by TLC in a solvent system consisting of chloroform/methanol/ammonia (65/35/5, v/v/v). Treatment of native erythrocyte membrane lipid micelles with phospholipase D, followed by reaction with diacetyl monoxime, suggests the presence of intrinsic carb-PE (2.85 +/- 0.65 percent of the total PE).
Carbamylation of erythrocyte aminophospholipid may be involved in some of the hematological consequences of uremia on the erythrocyte.
体外研究氰酸盐与红细胞膜氨基磷脂的结合情况，并探究天然红细胞质膜中是否能检测到氨甲酰化的氨基磷脂。
体外研究中，通过薄层层析（TLC）分离14C - 氨甲酰化红细胞膜的脂质成分。通过放射自显影观察氰酸盐的共价掺入情况，并通过磷分析进行定量。体内研究中，磷脂酶D酶解磷脂头部基团，随后与二乙酰单肟反应。
孵育15小时后，磷脂酰乙醇胺（PE）和磷脂酰丝氨酸（PS）均被[14C]氰酸盐共价修饰，分别掺入15.76±0.09和13.34±0.81摩尔％。在由氯仿/甲醇/氨（65/35/5，v/v/v）组成的溶剂系统中，通过TLC将氨甲酰化的PE（carb - PE）与PE分离。用磷脂酶D处理天然红细胞膜脂质微团，随后与二乙酰单肟反应，表明存在内源性carb - PE（占总PE的2.85±0.65％）。
红细胞氨基磷脂的氨甲酰化可能与尿毒症对红细胞产生的某些血液学后果有关。
