Belcourt A, Gillmeth S
Calcif Tissue Int. 1979 Nov 6;28(3):227-31. doi: 10.1007/BF02441240.
Pure human mature enamel was prepared using a careful microdissection technique. After EDTA dissolution, the soluble proteins were recovered representing a concentration of 0.035% in the initial enamel. When the samples were analyzed with polyacrylamide gel electrophoresis, Coomassie Brilliant Blue staining revealed only one sharp fast migrating band, whereas o-toluidine blue, methylene blue, Amido Black 10B, and pyronine red G showed a thin double band at the same migration distance. Ultracentrifugation studies suggested that the proteins were of low molecular weight or of weak density. Absorption spectra showed a strong absorbance at 260 nm. After hydrolysis, amino acid analyses yielded a composition of 25% Gly, 13.5% Glu, 11% Ser, and 11% Pro. Cysteine measured as cysteic acid was present at 2%, and 2% hydroxyproline was found. A carbohydrate content of 15% was estimated by the anthrone method. Glucose, galactose, mannose, and fucose, identified through gas chromatography, were in a molar ratio of 9:4:3:1. Thus the organic matrix of adult human enamel consists of one or possibly two acidic glycoproteins.
使用精细的显微解剖技术制备了纯人类成熟牙釉质。经乙二胺四乙酸(EDTA)溶解后,回收了可溶性蛋白质,其在初始牙釉质中的浓度为0.035%。当用聚丙烯酰胺凝胶电泳分析样品时,考马斯亮蓝染色仅显示出一条清晰的快速迁移带,而邻甲苯胺蓝、亚甲蓝、氨基黑10B和派洛宁红G在相同迁移距离处显示出一条细的双带。超速离心研究表明,这些蛋白质分子量低或密度小。吸收光谱显示在260nm处有强吸收。水解后,氨基酸分析得出的组成是:甘氨酸25%、谷氨酸13.5%、丝氨酸11%、脯氨酸11%。以半胱氨酸形式测得的半胱氨酸含量为2%,还发现了2%的羟脯氨酸。通过蒽酮法估计碳水化合物含量为15%。通过气相色谱鉴定的葡萄糖、半乳糖、甘露糖和岩藻糖的摩尔比为9:4:3:1。因此,成人牙釉质的有机基质由一种或可能两种酸性糖蛋白组成。