刘睿;蔡朔;赵珂璇;蒋梦彤;郑云枫;徐浩坤;侯蓉;黄勇;赵明;段金廒;
LIU Rui;CAI Shuo;ZHAO Ke-xuan;JIANG Meng-tong;ZHENG Yun-feng;XU Hao-kun;HOU Rong;HUANG Yong;ZHAO Ming;DUAN Jin-ao;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine;School of Pharmacy, Nanjing University of Chinese Medicine;Jiangsu Key Laboratory of Research and Development in Marine Bi

• 1:南京中医药大学中药资源产业化与方剂创新药物国家地方联合工程研究中心江苏省中药资源产业化过程协同创新中心
• 2:南京中医药大学药学院
• 3:南京中医药大学江苏省海洋药用生物资源研究与开发重点实验室
• 4:贵州广济堂健康药业有限公司

摘要(Abstract)：

该文采用纳升液相-串联质谱法对鹿皮胶样品进行分析,以PEAKS Studio蛋白质搜库鉴定软件对鹿皮胶赖氨酸位点的羟基化与O-糖基化修饰进行全面鉴定,明确鹿皮胶Ⅰ型胶原蛋白α1链(COL1A1)与α2链(COL1A2)发生羟基化与O-糖基化的主要位点、类型及数量。结果从4批次鹿皮胶样品中共鉴定了5 284个肽段信息,主要来源于COL1A1与COL1A2,其中羟基化赖氨酸(hydroxylysine, Hyl)共449个,半乳糖基化赖氨酸(galactosyl-hydroxylysine, Gal-Hyl)共442个,葡萄糖半乳糖基化赖氨酸(glucosyl-galactosyl-hydroxylysine, Glc-Gal-Hyl)共449个。鹿皮胶的Ⅰ型胶原发生羟基化与O-糖基化的主要位点有:N-末端肽的α1-9N与α2-5N,三螺旋结构域的α1-87,α1-174,α1-930,α2-87,α2-174,α2-933,C-末端肽的α1-16C。这些羟基化与糖基化赖氨酸与Ⅰ型胶原的三螺旋结构及胶原纤维形成密切相关,而鹿皮经高温高压熬制的过程破坏胶原的共价连接有利于胶原蛋白与胶原肽的溶出,形成具有糖基化修饰的胶原肽类成分。该文研究为鹿皮胶的功效物质基础的深入揭示提供了研究思路与方法依据,具有较好的理论意义与应用价值。
Nano-LC MS/MS was used to analyze trypsin digested deer-hide gelatin(DHG) samples, hydroxylation and O-glycosylation on lysine sites of DHG were comprehensive identified by using PEAKS Studio software. The sites, sorts and amounts of hydroxylation and O-glycosylation on Type Ⅰ collagen α1 chain(COL1 A1) and α2 chain(COL1 A2) of DHG were revealed. As a result, 5 284 peptides were identified from DHG samples, which were mainly from COL1 A1 and COL1 A2. Among these peptides, there were 449 peptides with hydroxylysine, 442 with galactosyl-hydroxylysine, 449 with glucosyl-galactosyl-hydroxylysine. The major modified sites of hydroxylation and O-glycosylation in DHG were shown as follow: α1-9 N and α2-5 N in N-telopeptides, α1-87, α1-174, α1-930, α2-87, α2-174, α2-933 in triple helix domain, and α1-16 C in C-telopeptides. These hydroxylation and O-glycosylation were correlated with the formation and stability of collagen molecules and collagen fibrils. It is feasible for the collagens and peptides dissolving from deer skin collagen fibrils under high temperature and pressure decocting, high temperature and pressure also might destroy inter-molecular covalent cross-linking and help those glycol-peptides formations. The present study provided ideas and strategies for the in-depth investigation on DHG chemical constituents, and showed good theoretical significance and application value.

关键词(KeyWords)： 鹿皮胶;修饰组;Ⅰ型胶原;赖氨酸位点;羟基化;糖基化
deer-hide gelatin;modifications;type Ⅰ collagen;lysine site;hydroxylation;glycosylation

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划项目(2018YFC1706100);; 国家自然科学基金面上项目(81973450);; 江苏省高校“青蓝工程”优秀青年骨干教师项目(2016年);; 江苏省“333工程”第三层次项目(2016年);; 贵州省科技攻关项目([2019]2771号,[2020]4Y090号);; 贵阳市科技局科技重大专项;; 中华中医药学会“青年人才托举工程”(QNRC2-C14);; 江苏省高等学校自然科学研究项目(19KJB360020)

作者(Author): 刘睿;蔡朔;赵珂璇;蒋梦彤;郑云枫;徐浩坤;侯蓉;黄勇;赵明;段金廒;
LIU Rui;CAI Shuo;ZHAO Ke-xuan;JIANG Meng-tong;ZHENG Yun-feng;XU Hao-kun;HOU Rong;HUANG Yong;ZHAO Ming;DUAN Jin-ao;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine;School of Pharmacy, Nanjing University of Chinese Medicine;Jiangsu Key Laboratory of Research and Development in Marine Bi

Email:

DOI: 10.19540/j.cnki.cjcmm.20201025.101

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