将牛关节软骨用NaCl和盐酸胍预处理,在酸性条件下用胃蛋白酶降解杂蛋白,经盐析、透析和冷冻干燥后获得Ⅱ型胶原,对其进行鉴定.结果表明,提取的Ⅱ型胶原纯度为92.2%,得率为61.5%,单条链分子量为130 k Da,氨基酸组成中Gly含量超过30%,Pro...将牛关节软骨用NaCl和盐酸胍预处理,在酸性条件下用胃蛋白酶降解杂蛋白,经盐析、透析和冷冻干燥后获得Ⅱ型胶原,对其进行鉴定.结果表明,提取的Ⅱ型胶原纯度为92.2%,得率为61.5%,单条链分子量为130 k Da,氨基酸组成中Gly含量超过30%,Pro与Hyp总含量超过20%,具有胶原特有的超分子结构,中性条件下变性温度为43.68℃,符合II型胶原特性.展开更多
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 p...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.展开更多
文摘将牛关节软骨用NaCl和盐酸胍预处理,在酸性条件下用胃蛋白酶降解杂蛋白,经盐析、透析和冷冻干燥后获得Ⅱ型胶原,对其进行鉴定.结果表明,提取的Ⅱ型胶原纯度为92.2%,得率为61.5%,单条链分子量为130 k Da,氨基酸组成中Gly含量超过30%,Pro与Hyp总含量超过20%,具有胶原特有的超分子结构,中性条件下变性温度为43.68℃,符合II型胶原特性.
基金Supported by National Natural Science Foundation of China (No.30970724)Natural Science Foundation of Tianjin (No.08JCYBJC03400)
文摘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.
