Identification of periodontal regeneration factor in poreine emanel protein

研究表明，釉质蛋白中含有牙周再生因子，已用于慢性牙周炎的治疗。然而，牙周再生因子与釉质蛋白的分离是必要的，因为牙周再生因子是由多种成分组成的，其中一些成分用于治疗时可能会对患者产生严重影响，从新形成的猪分泌性釉质中提取的釉质蛋白在狗下颌颊裂时具有牙周再生的活性，而从发育中的晚期分泌性釉质中提取的釉质蛋白则很少。用Sephadex G-100在0.05M碳酸盐缓冲液中进行凝胶过滤，将新形成的分泌性釉质中的釉质蛋白分离为4个组分。第一洗脱组分(Fr1)具有最强的牙周再生活性，含有89 kDa的釉蛋白、20~25 kDa的釉原蛋白和13~17 kDa的鞘蛋白。在这些组分中，经6M尿素离子交换层析和4M凝胶过滤层析分离的鞘蛋白组分具有牙周再生活性。然而，不可否认的是，鞘蛋白部分含有牙釉质蛋白中发现的骨形态发生蛋白或转化生长因子-β等生长因子。重组鞘蛋白是否具有牙周再生的活性是必须考察的。目前，我们尝试利用大肠杆菌连接系统来制备重组鞘蛋白

It is shown that enamel protein contains a periodontal regeneration factor and is already used for the therapy of chronic periodontitis. However, it is necessary to separate the periodontal regeneration factor from the enamel protein since it consists of multi-components of which some are possible to be affected severely to the patient when it is used for the therapy, The enamel protein extracted from the newly formed secretory enamel of pig had the activity for periodontal regeneration in a buccal dehiscence in dog mandible while that extracted from the advanced developing secretory enamel had scarce. The enamel protein of newly formed secretory enamel was separated into 4 fractions by the gel filtration using with a Sephadex G-100 in 0.05M carbonate buffer. The first eluted fraction (Fr 1) had the strongest activity for the periodontal regeneration in these fractions and contained the aggregate consisted of 89kDa enamelin 20〜25kDa amelogenins and 13〜17kDa sheath proteins. In these components in the aggregate the sheath protein fraction, which were separated by ion exchange chromatography in 6M urea solution and gel filtration chromatography in 4M guanidine solution was found to have the activity for periodontal regeneration. However, it is not denied that the sheath protein fraction contains the growth factors like BMP or TGF-β which are found in the enamel protein. It is necessarily examined whether r not recombinant sheath protein have the activity for periodontal regeneration.At present, we try to make the recombinant sheath protein using E coli ligation system