SYNTHESIS AND FUNCTION OF COMPOSITE SALIVARY MOLECULES

唾液复合分子的合成及功能

• 批准号: 3896864
• 负责人: MICHAEL J LEVINE
• 金额: --
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3896864
• 关键词: Streptococcus sanguis; adhesin; aluminum; amidation /deamidation; carbohydrates; chemical structure function; cysteine; glycopeptides; glycoproteins; human subject; hydroxyapatites; mucins; oligosaccharides; oral bacteria; phosphoproteins; proline; protein biosynthesis; protein glutamine gamma glutamyltransferase; saliva; sialate; transport proteins

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.

该子项目的总体目标是开始“新一代” 根据我们实验室获得的信息进行的研究 过去十年的结构/功能关系 几组唾液分子。 该信息有 允许我们为各种结构分配特定的功能 人类唾液粘蛋白​​（富含脯氨酸的糖蛋白）上的结构域 和含半胱氨酸的磷蛋白。 低聚糖 主要富含脯氨酸的糖蛋白（PRG）和低脯氨酸的单位 人唾液粘蛋白​​ (MG2) 的分子量已被证明 与唾液酸结合粘附素特异性相互作用 血链球菌表面并表现出润滑活性。 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 参与这些职能的机构仍然是本次会议的一个目标 子项目。 我们将利用现有信息开始准备工作 以及嵌合唾液分子的体外测试 双复合材料和三复合材料的性质。 双复合分子 将由 PRG 或 MG2 的碳水化合物单元组成 与完整的“载体”分子（例如白蛋白）共价偶联 和中性含半胱氨酸磷蛋白(CCP1)。 下一个， 我们将制备更先进的双复合分子系列 由白蛋白或CCP1的OHAp结合域组成 与 PRG 或 MG2 的碳水化合物单元共价偶联。 最后，我们将制备由以下组成的三复合分子： PRG 和 MG2 的碳水化合物单元与完整白蛋白偶联或 CCPl。碳水化合物单元（如糖肽）将偶联到 通过酶促（转谷氨酰胺酶）或化学作用的肽部分 （还原酰胺化）方法。 这将使我们能够准备一份 包含不同数量的分子群体 碳水化合物单位。 这个新糖蛋白群体将允许 我们检查碳水化合物的密度和数量的影响 介导链球菌相互作用和润滑的单位。 我们将采用的技术来准备 新糖蛋白包括复杂的表征 碳水化合物、自动肽测序和自动 肽合成技术。 所获得的信息对于以下方面可能至关重要 最终生产替代疗法（人工 唾液）用于患有唾液的个体的治疗用途 功能障碍或咬合磨损。 综上所述，本子项目 让我们有机会探索潜在价值 为患者设计的“复合或定制”唾液分子 个人需要。