| Extraction and Isolation of Type Ⅰ Ⅲ and Ⅴ Collagens and Their SDS-PAGE Analyses |
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| 引用本文： | 武继民,李志宏,袁晓燕,汪鹏飞,刘永清,王赫.Extraction and Isolation of Type Ⅰ Ⅲ and Ⅴ Collagens and Their SDS-PAGE Analyses[J].天津大学学报(英文版),2011,17(2):111-117. |
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| 作者姓名： | 武继民 李志宏 袁晓燕 汪鹏飞 刘永清 王赫 |
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| 作者单位： | Tianjin Key Laboratory of Composite and Functional Materials,School of Materials Science and Engineering,Tianjin University;Institute of Medical Equipment,Academy of Military Medical Sciences |
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| 基金项目： | Supported by National Natural Science Foundation of China (No.30970724);Natural Science Foundation of Tianjin (No.08JCYBJC03400) |
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| 摘 要： | Type Ⅰ,Ⅲ and Ⅴ collagens were extracted from bovine dermis and cornea by using pepsin treatment in acetic acid solution,followed by salt precipitation and dialysis,to purify and isolate each type of collagens.The preparation process was analyzed by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).A reducing agent,2-mercaptoethanol,was used to remove disulfide bonds and analyze the structure of the bonds involved between α chains in some types of collagens.The use of delayed reducing methods resulted in the difference between α1(Ⅲ) and α1(Ⅰ) chains in a mixture containing type Ⅰ and Ⅲ collagens.The structure of disulfide bonds among α chains exists potentially in type Ⅴ collagen prepared from the pepsin-treatment extraction at 4℃,which differs from type Ⅲ collagen in relation to the locations of disulfide bonds.Compared with pepsin-treated collagen at 4℃,the relative molecular weights of α1(Ⅴ) and α2(Ⅴ) chains treated at room temperature decrease by 4.6% and 6.0%,respectively.It is concluded that type Ⅰ,Ⅲ and Ⅴ collagens can be prepared from bovine dermis and cornea by the use of pepsin treatment,salt precipitation and dialysis.The interchain disulfide bonds lie potentially near the edges of termini of type Ⅴ collagen molecules in extracellular matrix,and a small number of interchain crosslinks exist in type Ⅴ collagen.
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| 关 键 词： | PAGE电泳 胶原蛋白 胃蛋白酶 SDS 提取 电泳分析 分离型 相对分子质量 |
Extraction and isolation of type I, III and V collagens and their SDS-PAGE analyses |
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| Jimin Wu, Zhihong Li, Xiaoyan Yuan, Pengfei Wang, Yongqing Liu and He Wang.Extraction and isolation of type I, III and V collagens and their SDS-PAGE analyses[J].Transactions of Tianjin University,2011,17(2):111-117. |
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| Authors: | Jimin Wu Zhihong Li Xiaoyan Yuan Pengfei Wang Yongqing Liu and He Wang |
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| Institution: | (1) Cancer Cellular Therapeutics, Inc., Cape Girardeau, MO, USA;(2) Hematology-Oncology Associates of Southeast Missouri Hospital, Southeast Medical Plaza, Cape Girardeau, MO, USA;; |
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| Abstract: | Type I, III and V collagens were extracted from bovine dermis and cornea by using pepsin treatment in acetic acid solution,
followed by salt precipitation and dialysis, to purify and isolate each type of collagens. The preparation process was analyzed
by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A reducing agent, 2-mercaptoethanol, was used
to remove disulfide bonds and analyze the structure of the bonds involved between α chains in some types of collagens. The use of delayed reducing methods resulted in the difference between α1(III) and α 1(I) chains in a mixture containing type I and III collagens. The structure of disulfide bonds among α chains exists potentially in type V collagen prepared from the pepsin-treatment extraction at 4 °C, which differs from type
III collagen in relation to the locations of disulfide bonds. Compared with pepsin-treated collagen at 4 °C, the relative
molecular weights of α1(V) and α2(V) chains treated at room temperature decrease by 4.6% and 6.0%, respectively. It is concluded that type I, III and V collagens
can be prepared from bovine dermis and cornea by the use of pepsin treatment, salt precipitation and dialysis. The interchain
disulfide bonds lie potentially near the edges of termini of type V collagen molecules in extracellular matrix, and a small
number of interchain crosslinks exist in type V collagen. |
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| Keywords: | collagen interchain SDS-PAGE preparation structure |
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