ISOLATION AND CHARACTERIZATION OF A DISTINCT TYPE OF COLLAGEN FROM BOVINE FETAL MEMBRANES AND OTHER TISSUES
ISOLATION AND CHARACTERIZATION OF A DISTINCT TYPE OF COLLAGEN FROM BOVINE FETAL MEMBRANES AND OTHER TISSUES
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DOI：
10.1021/bi00587a003
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发表时间：
1979-01-01
期刊：
影响因子：
2.9
通讯作者：
CANNON, DJ
中科院分区：
文献类型：
--
作者：
HONG, BS;DAVISON, PF;CANNON, DJ
开发了一种新的方法来分离含有αA和αB链的独特的牛胶原蛋白部分。该组分首先从牛羊膜和绒毛膜中获得，然后将该方法应用于从角膜、肌腱、皮肤、硬脑膜、神经内膜和软骨中溶解的胃酶增溶胶原蛋白。透射电子显微镜显示单一类型的长间距节段微晶，染色特征与以前在牛后叶S膜和心脏瓣膜的胶原蛋白中区分的VI型节段相同。αA和αB链在羧甲基纤维素上是不可分离的，但可通过羟基磷灰石层析进行拆分。αA与αB的链比为1：1.78。氨基酸分析表明，αA和αB链的组成是可区分的，并类似于发表的对人胶原蛋白同源链的分析。没有检测到半胱氨酸或二硫键。糖残基的两条链的碱性水解物分析表明，αB中84%的羟基赖氨酸和αA中45%的羟赖氨酸是糖基化的。天然胶原蛋白对哺乳动物胶原酶有明显的抵抗力。αA和αB链显然都存在于天然αA(αB)2分子中，因为当从各种组织中分离时，该胶原蛋白始终以大约1：2的比例包含αA和αB链，在这些制剂中EM只能检测到一种类型的链段长间隔聚集体，并且这两条链都对胶原酶具有抗性。
A new procedure was developed to isolate a distinct bovine collagen fraction which contains .alpha.A and .alpha.B chains. This fraction was obtained first from the bovine amnion and chorion, and the method was subsequently applied to the pepsin-solubilized collagens dissolved from cornea, tendon, skin, dura, nerve endoneurium and cartilage. EM reveals a single type of segment long-spacing crystallite, identical in staining characteristics with the type VI segments previously distinguished among the collagens in bovine Descemet''s membrane and heart valve. The .alpha.A and .alpha.B chains were inseparable on carboxymethylcellulose but were resolved by hydroxyapatite chromatography. The .alpha.A to .alpha.B chain ratio was 1:1.78. Amino acid analysis demonstrated that the compositions of the .alpha.A and .alpha.B chains are distinguishable and similar to analyses published for the homologous chains from human collagen. No cysteine or disulfide linkages were detected. Alkaline hydrolysate analysis of both chains for sugar residues indicated that 84% of the hydroxylysines in .alpha.B and 45% in .alpha.A are glycosylated. The native collagen is apparently resistant to mammalian collagenase. The .alpha.A and .alpha.B chains are evidently both present in a native .alpha.A(.alpha.B)2 molecule because when isolated from various tissues this collagen consistently contains the .alpha.A and .alpha.B chains in a ratio approximating 1:2, only a single type of segment long-spacing aggregate could be detected by EM in these preparations and both chains are resistant to collagenase.