SALIVARY PROTEINS--MOLECULAR STUDY OF STRUCTURE-FUNCTION

唾液蛋白——结构功能的分子研究

• 批准号: 3223540
• 负责人: LIBUSE ANNA BOBEK
• 金额: $ 11.7万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-03-15 至 1997-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3223540
• 关键词: Candida albicans; Escherichia coli; antibacterial agents; antifungal agents; antiinfective agents; antiviral agents; chimeric proteins; chromatography; circular dichroism; cysteine; gene deletion mutation; molecular biology; molecular cloning; molecular site; mutant; oligonucleotides; papain; polymerase chain reaction; protease inhibitor; protein purification; protein structure; protein structure function; recombinant DNA; saliva; site directed mutagenesis

Molecules found in human saliva have many protective functions critical to oral health. The protective qualities of saliva become evident in individuals with markedly decreased salivary flow (xerostomia or dry mouth). The long range goal of our research is to design and produce the individual and/or chimeric human salivary molecules with enhanced protective functions and to use these as prototypes for development of artificial salivas. An essential prerequisite for this goal is the complete understanding of the structure-function relationship of individual salivary molecules. The availability of mutants provides one of the most powerful ways to achieve this objective. The use of molecular biological techniques and recombinant DNA technology proposed in this application for the production the native, variant (mutant) and chimeric forms of proteins in the heterologous system (E coli) will allow us to study the salivary protein's structure and their function at the molecular level and permit a rational design of salivary substitutes. Recently, we have obtained recombinant clones encoding several types of salivary proteins which will enable us to carry out the proposed studies. Molecules that we will concentrate on are cystatins (cysteine proteinase inhibitors and anti-bacterial/viral agents) and histatins (anti-fungal, particularly anti-candidal and anti-bacterial agents), since they have key protective functions and are amongst the best characterized salivary molecules on a biochemical basis. Specifically, we propose to: 1)produce large amounts of biologically active cystatin and histatin in E coli expression system, 2) construct mutants of cystatin and histatin which will be used for mapping the structural domains and/or individual amino acid residues that are essential for the structural integrity and/or biological function of these proteins and 3) construct and produce various cystatin/histatin chimeric proteins with biological activities resembling or exceeding those of the parent proteins.

在人类唾液中发现的分子具有许多保护功能关键 口腔健康。 唾液的保护特质在 唾液流量显着降低的人（静脉或干燥 嘴）。 我们研究的远距离目标是设计和生产 具有增强的个体和/或嵌合人唾液分子 保护功能并将其用作开发的原型 人造唾液。 这个目标的基本先决条件是 完全了解结构功能关系的关系 个体唾液分子。 突变体的可用性提供了一个 实现这一目标的最有力方法。 使用 提出的分子生物学技术和重组DNA技术 在本生产申请中，天然，变体（突变体）和 异源系统（E Coli）中蛋白质的嵌合形式将 允许我们研究唾液蛋白的结构及其功能 分子水平并允许唾液的合理设计 替代品。 最近，我们获得了编码的重组克隆 几种类型的唾液蛋白将使我们能够执行 拟议的研究。 我们将集中精力的分子是囊蛋白 （半胱氨酸蛋白酶抑制剂和抗细菌/病毒剂）和 组织蛋白（抗真菌，尤其是抗菌和抗细菌 代理），因为它们具有关键的保护功能，并且是 在生化的基础上，最能表征唾液分子。 具体而言，我们提出：1）生物学大量生物 活性胱抑素和组蛋白在E大肠杆菌表达系统中，2）构造 胱抑素和组织蛋白的突变体，用于映射 结构域和/或单个氨基酸残基 对于结构完整性和/或生物学功能至关重要 这些蛋白质和3）构建并产生各种胱抑素/组蛋白 具有类似或超过生物活性的嵌合蛋白 父蛋白的那些。