N-EPSILON-(CARBOXYMETHYL)LYSINE IS A DOMINANT ADVANCED GLYCATION END-PRODUCT (AGE) ANTIGEN IN TISSUE PROTEINS
N-EPSILON-(CARBOXYMETHYL)LYSINE IS A DOMINANT ADVANCED GLYCATION END-PRODUCT (AGE) ANTIGEN IN TISSUE PROTEINS
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DOI：
10.1021/bi00034a021
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发表时间：
1995-08-29
期刊：
影响因子：
2.9
通讯作者：
BAYNES, JW
中科院分区：
文献类型：
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作者：
REDDY, S;BICHLER, J;BAYNES, JW
糖基化终末产物（AGEs）和糖氧化产物是糖和蛋白质之间的美拉德或布朗宁反应过程中形成的，并且与衰老的病理生理学和糖尿病的并发症有关。为了确定AGEs的结构，制备了通过与葡萄糖孵育而褐变的蛋白质的抗体，并用于ELISA测定，以测量在牛血清白蛋白（BSA）或N-α-乙酰赖氨酸与葡萄糖、果糖或乙二醛之间的模型反应中形成的AGEs。糖基化终产物仅在氧化条件下由葡萄糖和果糖形成，但在氧化和抗氧化条件下由乙二醛形成。凝胶渗透色谱分析表明，在N-α-乙酰基赖氨酸与葡萄糖、果糖和乙二醛的反应中形成了类似的AGE，并且该AGE与真实的N-α-乙酰基-N-α-（羧甲基）赖氨酸共洗脱。AGE蛋白的氨基酸分析显示N-羧甲基赖氨酸（CML）的含量显着。在使用针对AGE蛋白的多克隆抗体的ELISA测定中，通过BSA的化学修饰制备的CML-BSA（类似于25 mol CML/mol BSA）是AGE蛋白和人透镜蛋白中AGE的识别的有效抑制剂。我们的结论是，AGEs主要是糖氧化产物，CML是一个主要的AGE组织蛋白中的AGE蛋白的多克隆抗体。
Advanced glycation end products (AGEs) and glycoxidation products are formed during Maillard or browning reactions between sugars and proteins and are implicated in the pathophysiology of aging and the complications of diabetes. To determine the structure of AGEs, antibodies were prepared to protein browned by incubation with glucose and used in ELISA assays to measure AGEs formed in model reactions between bovine serum albumin (BSA) or N-alpha-acetyllysine and glucose, fructose, or glyoxal. AGEs were formed from glucose and fructose only under oxidative conditions, but from glyoxal under both oxidative and antioxidative conditions. Gel permeation chromatographic analysis indicated that a similar AGE was formed in reactions of N-alpha-acetyllysine with glucose, fructose, and glyoxal and that this AGE co-eluted with authentic N-alpha-acetyl-N-epsilon-(carboxymethyl)lysine. Amino acid analysis of AGE proteins revealed a significant content of N-epsilon-(carboxymethyl)lysine (CML). In ELISA assays using polyclonal antibodies against AGE proteins, CML-BSA (similar to 25 mol of CML/mol of BSA), prepared by chemical modification of BSA, was a potent inhibitor of the recognition of AGE proteins and of AGEs in human lens proteins. We conclude that AGEs are largely glycoxidation products and that CML is a major AGE recognized in tissue proteins by polyclonal antibodies to AGE proteins.