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为了分离纯化骨素及其酶解产物中的蛋白质及肽,并比较骨素蛋白与骨蛋白和肌肉蛋白的差异,本文以骨蛋白和肌肉蛋白为参照,采用十二烷基磺酸钠凝胶电泳法(SDS-PAGE)对骨素蛋白及其酶解产物中的肽进行分离。利用不同浓度的分离胶(5%、7.5%、10%、15%)对骨素蛋白、骨蛋白和肌肉蛋白样品进行分离,结果发现,各种浓度的分离胶均可以分离骨蛋白和肌肉蛋白,但是不能分离骨素蛋白。对骨素蛋白酶解液采用N-三(羟甲基)甲基甘氨酸凝胶电泳法分离,能有效分离骨素酶解液中的肽。对SDS-PAGE电泳分离时骨蛋白的迁移比和分离胶的相关性分析发现,骨蛋白的迁移比与分离胶浓度间呈指数相关,且具有极显著的相关性(R2≥0.996,p<0.01)。利用该相关性并结合蛋白质的分子量与迁移比的相关性方程,可以通过蛋白质的分子量选择合理的分离胶浓度,或者通过迁移比预测蛋白质的分子量。
In order to separate and purify the proteins and peptides in bone and its hydrolyzate, and to compare the difference between osteoprotegerin and bone protein and muscle protein, we used sodium dodecyl sulfate gel Electrophoresis (SDS-PAGE) separates the peptides from the osteoprotegerin and its hydrolysates. Using different concentrations of separation gums (5%, 7.5%, 10%, 15%) for the separation of osteopontin, osteopontin and muscle protein samples, it was found that various concentrations of separation gel were able to separate bone proteins and muscle proteins , But can not separate osteoprotein. The separation of the osteoprotegerin from N-tris (hydroxymethyl) methylglycine gel electrophoresis can effectively separate the peptides in the digestase solution. The correlation between the migration ratio of bone protein and the separation gel during the SDS-PAGE electrophoresis separation showed that there was an exponential correlation between the migration ratio of the bone protein and the concentration of the separation gel, and had a very significant correlation (R2≥0.996, p <0.01 ). Using this correlation and in combination with the equation of correlation of the molecular weight and the migration ratio of the protein, a reasonable separation gel concentration can be selected by the molecular weight of the protein, or the molecular weight of the protein can be predicted by the migration ratio.