Ramshaw J A, Werkmeister J A, Bremner H A
CSIRO Division of Protein Chemistry, Parkville VIC, Australia.
Arch Biochem Biophys. 1988 Dec;267(2):497-502. doi: 10.1016/0003-9861(88)90056-2.
The skin collagen of a fish, blue grenadier (Macruronus novaezelandiae), has been purified and characterized. The fish skin was readily soluble in dilute acetic acid, with no pepsin treatment needed. The collagen was purified by salt precipitation. Skin samples from fish of various ages showed that even in the oldest sample, more than 8 years of age, the collagen was still readily acid soluble. The purified collagen had a melting temperature of 22 degrees C; the shrinkage temperature for the skin was 48 degrees C. Its tissue distribution, examined by immunohistology, and its chemical properties indicated a close homology to mammalian type I collagen. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that three distinct alpha-chains were present. These were purified by ion-exchange chromatography on CM-cellulose and by gel permeation chromatography on Superose 6. The three purified alpha-chain fractions were examined by amino acid analysis and by SDS-PAGE of their cyanogen bromide fragments. These data indicated that the additional chain was genetically distinct, and most closely related to the alpha 1-chain, from which it was poorly resolved on SDS-PAGE.
新西兰无须鳕（Macruronus novaezelandiae）鱼皮中的胶原蛋白已被纯化并进行了特性分析。鱼皮很容易溶解于稀醋酸中，无需胃蛋白酶处理。通过盐沉淀法对胶原蛋白进行了纯化。来自不同年龄鱼的皮肤样本表明，即使是年龄超过8岁的最老样本中的胶原蛋白仍很容易酸溶。纯化后的胶原蛋白的熔点为22℃；鱼皮的收缩温度为48℃。通过免疫组织学检查其组织分布及其化学性质表明，它与哺乳动物I型胶原具有密切的同源性。然而，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）显示存在三条不同的α链。通过在CM-纤维素上进行离子交换色谱和在Superose 6上进行凝胶渗透色谱对它们进行了纯化。通过氨基酸分析及其溴化氰片段的SDS-PAGE对三个纯化的α链组分进行了检测。这些数据表明，额外的链在遗传上是不同的，并且与α1链关系最为密切，在SDS-PAGE上它与α1链很难区分开来。
