Use of 1H NMR ROESY for structural determination of O-glycosylated amino acids from a serine-containing glycopeptidolipid antigen.
Use of 1H NMR ROESY for structural determination of O-glycosylated amino acids from a serine-containing glycopeptidolipid antigen.
复制标题
使用 1 H NMR ROESY 对含丝氨酸糖肽脂抗原中的 O-糖基化氨基酸进行结构测定。
DOI：
10.1021/bi00129a004
复制
发表时间：
1992
期刊：
影响因子：
2.9
通讯作者：
G. Puzo
中科院分区：
文献类型：
--
作者：
M. Rivière;G. Puzo
从蟾蜍分枝杆菌中分离出的含有丝氨酸的糖肽脂抗原代表了一类新的分枝杆菌糖肽脂抗原，其缺乏C-菌苷核心结构[Rivière, M., & Puzo, G. (1991) J. Biol.。化学。 266、9057-9063]。分配给 C12-Ser-Ser-Phe-alloThr-OCH3 的脂肽核心表现出三个潜在的糖基化位点。碳水化合物部分由 3-O-甲基-6-脱氧-α-L-吡喃糖基和 2,3,4-三-O-甲基-L-吡喃鼠李糖基(α 1----3)-2-O-月桂酰-L-吡喃鼠李糖基(α 1----3)-L-吡喃鼠李糖基(α 1----3)-2,4-二-O-(乙酰基，月桂酰)-6-脱氧-α-L-吡喃葡萄糖基附属物。在目前的工作中，通过使用氯仿作为溶剂对天然糖肽脂进行 ROESY 实验，成功地确定了碳水化合物的附着位点。从NOE接触中，我们明确地确定酰化丝氨酸被3-O-甲基-6-脱氧-α-L-吡喃鼠糖基附属物糖基化，而2,3,4-三-O-甲基-L-吡喃鼠李糖基(α 1----3)-2-O-月桂酰-L-吡喃鼠李糖基(α 1----3)-L-吡喃鼠李糖基(α 1----3)-2,4-二-O-(乙酰基，月桂酰基)-6-脱氧-α-L-吡喃葡萄糖基附属物与C端异位Thr-OCH3结合。根据这些数据，成功定位了基础单糖单元的 C-2 和 C-4 上的乙酰基和月桂酰基残基。此外，建立了丝氨酸和苯丙氨酸残基的“L”绝对构型以及异苏氨酸的“D”构型。这种新型分枝杆菌抗原（一种含丝氨酸的糖肽脂）的一级结构现已完全确定。
A serine-containing glycopeptidolipid antigen isolated from Mycobacterium xenopi typified a new class of mycobacterial glycopeptidolipid antigens devoid of the C-mycoside core structure [Rivière, M., & Puzo, G. (1991) J. Biol. Chem. 266, 9057-9063]. The lipopeptide core assigned to C12-Ser-Ser-Phe-alloThr-OCH3 exhibits three potential sites of glycosylation. The carbohydrate parts are composed of 3-O-methyl-6-deoxy-alpha-L-talopyranosyl and 2,3,4-tri-O-methyl-L- rhamnopyranosyl(alpha 1----3)-2-O-lauroyl-L-rhamnopyranosyl(alpha 1----3)-L- rhamnopyranosyl(alpha 1----3)-2,4-di-O-(acetyl, lauroyl)-6-deoxy-alpha-L-glucopyranosyl appendages. In the present work, the carbohydrate attachment sites were successfully determined by ROESY experiments on the native glycopeptidolipid using chloroform as solvent. From the NOE contacts, we unambiguously established that the acylated serine is glycosylated by the 3-O-methyl-6-deoxy-alpha-L-talopyranosyl appendage while the 2,3,4-tri-O-methyl-L-rhamnopyranosyl(alpha 1----3)-2-O- lauroyl-L-rhamnopyranosyl(alpha 1----3)-L-rhamnopyranosyl(alpha 1----3)-2,4-di- O-(acetyl, lauroyl)-6-deoxy-alpha-L-glucopyranosyl appendage is bound to the C-terminal alloThr-OCH3. From these data, the acetyl and lauroyl residues on the C-2 and C-4 of the basal monosaccharide unit were successfully localized. Furthermore, the "L" absolute configuration for the serines and the phenylalanine residues and the "D" configuration for the allothreonine were established. The primary structure of this novel type of mycobacterial antigen, a serine-containing glycopeptidolipid, has now been fully established.