SYNTHESIS AND FUNCTION OF COMPOSITE SALIVARY MOLECULES

这个子项目的总体目标是开始“新一代” 基于我们实验室获得的信息 在过去的十年里， 几组唾液分子。 这一信息 允许我们为各种结构分配特定的功能， 富含脯氨酸的糖蛋白， 和含半胱氨酸的磷蛋白。 寡糖 单位的主要脯氨酸丰富的糖蛋白（PRG）和低 分子量的人唾液粘蛋白（MG 2）已被证明 特异性地与唾液酸结合粘附素相互作用， 具有润滑活性。 含半胱氨酸的磷蛋白（CCP's）已被证明 与羟基磷灰石（OHAp）结合并调节矿化 流程. 然而，CCP的精确结构域 参与这些功能的人仍然是这一目标的一部分。 子项目。 我们将利用我们现有的信息开始准备 和嵌合唾液分子的体外测试。 双复合和三复合性质。 双复合分子 将由PRG或MG 2的碳水化合物单元组成 与完整的“载体”分子如白蛋白共价偶联 和中性含半胱氨酸磷蛋白（CCP 1）。 下一个是 我们将准备一系列更先进的双复合分子 由白蛋白或CCP 1的OHAp结合结构域组成 在一些实施方案中，所述化合物与PRG或MG 2的碳水化合物单元共价偶联。 最后，我们将制备由 与完整白蛋白偶联的PRG和MG 2的碳水化合物单元，或 CCP 1.碳水化合物单元（如糖肽）将偶联至 通过酶促（转氨酶）或化学 （还原酰胺化）方法。 这将使我们能够准备一个 一群分子，其中含有不同数量的 碳水化合物单位 这群新糖蛋白将允许 我们来研究碳水化合物的密度和数量的影响 单位介导链球菌的相互作用和润滑。 我们将采用的技术 新糖蛋白包括复合物的表征 碳水化合物，自动化肽测序， 肽合成技术。 所获得的信息可能是至关重要的， 替代疗法的最终生产（人工 唾液）用于患有唾液腺疾病的个体的治疗用途 功能障碍或咬合磨损。 总而言之，本次项目 让我们有机会探索 “复合或定制”唾液分子设计用于患者的 个人需要。

The overall goal of this subproject is to begin a "new generation" of studies based upon information which our laboratory has gained over the last decade on the structure/function relationships of several groups of salivary molecules. This information has permitted us to assign specific functions to various structural domains on human salivary mucins, the proline-rich glycoprotein and cysteine-containing phosphoproteins. The oligosaccharide units of the major proline-rich glycoprotein (PRG) and the low molecular weight human salivary mucin (MG2) have been shown to specifically interact with sialic acid binding adhesins on the surface of streptococcus sanguis and exhibit lubricating activity. The cysteine-containing phosphoproteins (CCP's) have been shown to bind to hydroxyapatite (OHAp) and modulate mineralization processes. However, the precise structural domain(s) of CCPs which participates in these functions remains an objective of this subproject. We will utilize our current information to begin the preparation and in vitro testing of chimeric salivary molecules of a bicomposite and a tricomposite nature. Bicomposite molecules will consist of the carbohydrate units of the PRG or MG2 covalently coupled to intact "carrier" molecules such as albumin and the neutral cysteine-containing phosphoprotein (CCPl). Next, we will prepare a more advanced series of bicomposite molecules comprised of the OHAp binding domains of albumin or CCPl covalently coupled to the carbohydrate units of PRG or MG2. Finally, we will prepare tricomposite molecules comprised of the carbohydrate units of PRG and MG2 coupled to intact albumin or CCPl. Carbohydrate units (as glycopeptides) will be coupled to peptide moieties by enzymatic (transglutaminase) or chemical (reductive amidation) methods. This will enable us to prepare a population of molecules which contains a varying number of carbohydrate units. This population of neoglycoproteins will allow us to examine the effect of density and number of carbohydrate units in mediating streptococcal interactions and lubrication. The technologies which we will employ to prepare neoglycoproteins include characterization of complex carbohydrates, automated peptide sequencing, and automated peptide synthesis techniques. The information obtained could be of paramount importance in the eventual production of replacement therapies (artificial salivas) for therapeutic use by individuals with salivary dysfunction or occlusal abrasion. In summary, this subproject gives us the opportunity to explore the potential value of "composite or custom" salivary molecules designed for a patient's individual need.