摘要
选择ＳＤ大鼠切牙分离制备未成熟的牙釉质组织。参照Ｔｅｒｍｉｎｅ等的方法，采用盐酸胍及盐酸胍－ＥＤＴＡ分步提取法将牙釉质蛋白分为釉原蛋白和釉蛋白两大组分。经Ｓｅｐｈａｄｅｘ－Ｇ２００层析，初步将釉原蛋白分为Ａ１，Ａ２，Ａ３三个组分，将釉蛋白分为Ｅ１，Ｅ２两个组分。经ＳＤＳ－ＰＡＧＥ分析，所纯化之釉原蛋白Ａ１以２３ｋＤ为主要成分。在釉蛋白的层析主要分离组分Ｅ１中，蛋白含量极低，ＳＤＳ－ＰＡＧＥ测得其为４４ｋＤ的单一组分。釉质蛋白质在特定ｐＨ值，离子强度及溶液蛋白浓度改变后出现自我聚集沉淀。ＰＡＧＥ电泳分析提示，釉原蛋白分子中可能含有较多亚基。
Sprague Dawley rats were chosen to prepare immature and soft enamel tissue. The authors used a sequential dissociative extraction scheme in which the enamel matrix proteins were extracted first in guanidine HCl and then in guanidine HClEDTA, just as the method described by Termine, et al. The first step completely extracted amelogenins and the second step got enamelin. These two groups of protein were further purified by SephadexG200 gel filtration chromatography and analysed by polyacrylamide gel electrophoresis. The results showed: amelogenins were resolved into three fractions eluted from the column, and expressed as A1, A2, A3, respectively. The main fraction A1, which had a molecular weight range from 18 kD to 29 kD, had a main protein band of 23 kD when estimated by SDSPAGE. Enamelin was eluted into two fractions called E1 and E2. The main fraction E1 had a single band of 44 kD on the SDSPAGE gel. PAGE showed that amelogenins had many subunits. The authors also observed that the amelogenins would aggregate and precipitate reversiblly when pH value, ionic strenghth or protein concentration changed.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
1998年第1期12-14,T001,共4页
West China Journal of Stomatology
关键词
釉原蛋白
釉蛋白
大鼠
牙釉质
\ amelogenins\ \ enamelin\ \ enamel\ \ rat
