Characterization of bovine serum albumin glycated with glucose, galactose and lactose
Characterization of bovine serum albumin glycated with glucose, galactose and lactose
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DOI：
10.18388/abp.2008_3054
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发表时间：
2008-01-01
影响因子：
1.7
通讯作者：
Vazquez-Moreno, Luz
中科院分区：
文献类型：
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作者：
Irene Ledesma-Osuna, Ana;Ramos-Clamont, Gabriela;Vazquez-Moreno, Luz
还原糖和蛋白质之间的非酶促反应，称为糖化，已经受到营养和医学研究的越来越多的关注。此外，人们对获得用于生物学研究的纯的充分表征的寡糖的糖缀合物有很大的兴趣。在这项研究中，糖化牛血清白蛋白（BSA）的D-葡萄糖，D-半乳糖和D-乳糖在干热下在60 ℃ 30，60，120，180或240分钟进行了评估和糖化产物的研究，以建立其生物识别凝集素。使用凝胶电泳、可用氨基的测定和凝集素结合测定来监测BSA糖化。通过质谱和胰蛋白酶和糜蛋白酶消化研究BSA分子量增加和糖基化位点。取决于时间和糖的类型，实现BSA缀合的差异。SDS/PAGE显示修饰后BSA的氨基利用率降低，迁移速度减慢。D-半乳糖比D-葡萄糖或D-乳糖更具反应性，在反应120分钟后分别导致10、3和1个糖残基的偶联。BSA赖氨酸（K）是优选的修饰氨基酸; K256和K420似乎最可用于缀合。只有BSA-乳糖表现出特异性凝集素的生物识别。
The non-enzymatic reaction between reducing sugars and proteins, known as glycation, has received increased attention from nutritional and medical research. In addition, there is a large interest in obtaining glycoconjugates of pure well-characterized oligosaccharides for biological research. In this study, glycation of bovine serum albumin (BSA) by D-glucose, D-galactose and D-lactose under dry-heat at 60 degrees C for 30, 60, 120, 180 or 240 min was assessed and the glycated products studied in order to establish their biological recognition by lectins. BSA glycation was monitored using gel electrophoresis, determination of available amino groups and lectin binding assays. The BSA molecular mass increase and glycation sites were investigated by mass spectrometry and through digestion with trypsin and chymotrypsin. Depending on time and type of sugar, differences in BSA conjugation were achieved. Modified BSA revealed reduction of amino groups' availability and slower migration through SDS/PAGE. D-Galactose was more reactive than D-glucose or D-lactose, leading to the coupling of 10, 3 and 1 sugar residues, respectively, after 120 minutes of reaction. BSA lysines (K) were the preferred modified amino acids; both K256 and K420 appeared the most available for conjugation. Only BSA-lactose showed biological recognition by specific lectins.