驴真皮中主要蛋白的组成及其相互作用的研究

Major constituent proteins in donkey hide and their interaction

目的:分析阿胶主要原料驴真皮中的主要构成蛋白,研究其相互作用,揭示其分子构成,为阐明阿胶作用机制的研究提供实验依据。方法:分别使用TriPure试剂和通过1%聚丙烯酰胺(SDS)煮沸驴的真皮,提取驴皮中总蛋白质,再使用聚丙烯酰胺凝胶电泳分离蛋白质,割胶提取蛋白质,使用毛细管色谱进一步分析蛋白组分。结果:发现驴真皮中主要蛋白质不仅含有胶原蛋白α1(Ⅰ)型、α2(Ⅰ)型,而且还含有血清白蛋白,其中,用TriPure试剂提取蛋白质的总蛋白中,血清白蛋白的含量超过25%;用1%SDS煮沸的总蛋白中,血清白蛋白的含量约20%,并且发现在凝胶电泳带中有2条明显的分子质量为20万左右的蛋白质,割胶提取该蛋白质经过毛细管色谱分析,发现是胶原蛋白与血清白蛋白结合的产物。结论:血清白蛋白是驴真皮中的主要蛋白组分,驴血清白蛋白有可能对于阿胶补血的功能起到比普遍认为胶原蛋白更为重要的作用。

Objective： To analyze the constituent proteins in donkey hide, the key ingredient for Ejiao, an important traditional Chinese medicine for the blood-related conditions, in hope to eventually decipher the biochemical mechanism behind Ejiao＇s prominent medicinal efficacy. Method： Two methods were employed to extract pmtoins in donkey skin. One used TriPure isolation reagent to extract the total proreins in donkey skin. Anether used 1% sodium dodecyl sulfate（SDS） to heat the sample at 100℃ overnight. And then sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE） and capillary HPLC were used to analyze the component of proteins. Result： There are not only collagen al（ Ⅰ ） and collagen α2（ Ⅰ ）, but also serum albumin in donkey skin. The content is over 25% in total proteins with the method of TfiPure isolation reagent. The content of donkey serum albumin is up to 20% with the method of 1% SDS heating. And two bands,molecular weight are nearly 200 kDa,were found on 7.5% SDS-PAGE. Extracted these proteins to analyze with capillary HPLC, they were found to be the complex products of collagen and serum albumin of donkey. Conclusion： Donkey serum albumin is a main protein component in the hide, which is a clue to expose the effect of Ejiao on blood.