Oral Fluid Proteolytic Effects on Salivary Protein Structure and Function

口腔液蛋白水解对唾液蛋白质结构和功能的影响

• 批准号: 7595090
• 负责人: Eva Josephine Helmerhorst
• 金额: $ 20.31万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595090
• 关键词: Accounting; Address; Affect; Anti-Bacterial Agents; Antifungal Agents; Area; Biochemistry; Biological; Biological Assay; Biological Process; Body Fluids; Candida albicans; Characteristics; Dental Enamel; Dental caries; Disease; Duct (organ) structure; Environment; Enzymes; Etiology; Growth; Health; Homeostasis; Individual; Investigation; Liquid substance; Maintenance; Mass Spectrum Analysis; Minerals; Modeling; Oral; Oral cavity; Oral health; Organism; Parotid Gland; Peptide Hydrolases; Peptides; Periodontal Diseases; Physiological; Probability; Process; Property; Protein Family; Proteins; Proteolysis; Proteolytic Processing; Research; Role; Saliva; Salivary; Salivary Proteins; Source; Streptococcus mutans; Structure; Sublingual Gland; Submandibular gland; Surface; System; Testing; Work; antimicrobial; base; calcium phosphate precipitation; clinically significant; histatin 1; histatin 3; histidine-rich proteins; insight; macromolecule; novel; oral infection; proline-rich proteins; protein degradation; protein function; protein profiling; protein structure function; saliva function; statherin; translational approach; virtual

DESCRIPTION (provided by applicant): All biological processes occurring on oral surfaces in health and disease are dictated by the characteristics of the oral fluid environment. The major contributors to whole saliva are exocrine secretions derived from parotid, submandibular and sublingual glands. The predominant salivary proteins in these secretions should be readily identifiable in whole saliva. Surprisingly however, the protein profiles of glandular secretions differ significantly from that of whole saliva. Structure/function studies of salivary proteins isolated from glandular secretions have provided much insight into their potential roles in the maintenance of oral health. The drastic discrepancy between whole saliva and glandular secretions, however, has hardly been addressed in terms of its potential functional implications. One of the major differences between glandular secretions and whole saliva is the pronounced proteolytic activity of whole saliva. This activity appears to be in large part responsible for the apparent loss of intact proteins from this body fluid. Proline-rich proteins (PRPs), statherin and histatins have been well characterized in terms of their properties relating to mineral homeostasis and antimicrobial activity. While PRPs, statherin and histatins have been considered to be functionally among the most relevant salivary proteins, they also belong to those salivary proteins that are highly susceptible to proteolysis upon release into the oral cavity. Our hypothesis is that structural changes in proteins which occur between the release of secretion from glandular excretory ducts and the mixing of these proteins with the liquid and cellular constituents present in the oral cavity will have important functional, and hence physiological, consequences. This proposal focuses on the impact of salivary proteolysis on the structure and function of PRP-1, statherin, histatin-1 and histatin-3 by: 1. Studying the rate and mode of salivary protein degradation using as enzyme sources both liquid and cellular fractions of whole saliva. The degradation products will be subjected to chromatographic separation by RPHPLC and the individual peptides will be identified and characterized by mass spectrometry. 2. Determining the effect of whole saliva proteolysis on protein functions that are directly related to oral health. To achieve this, protein degradation mixtures and individual fragments will be evaluated in assays assessing their capacity to inhibit primary and secondary calcium phosphate precipitation as well as antifungal and antibacterial activities. Project Narrative: To sustain oral health, proper saliva function is a prerequisite. The current proposal will investigate how salivary proteolytic enzymes affect the structure and function of four prominent and biologically important glandular salivary proteins.

描述（由申请人提供）：健康和疾病中口腔表面上发生的所有生物过程均取决于口服液体环境的特征。整个唾液的主要因素是源自腮腺，下颌和舌下腺的外分泌分泌物。这些分泌物中的主要唾液蛋白应在整个唾液中很容易鉴定。然而，令人惊讶的是，腺分泌物的蛋白质谱与整个唾液的蛋白质谱图显着不同。从腺体分泌物分离的唾液蛋白的结构/功能研究为它们在维持口腔健康中的潜在作用提供了很多深入的了解。然而，整个唾液和腺体分泌之间的巨大差异几乎没有得到其潜在功能的影响。腺体分泌物和整个唾液之间的主要差异之一是整个唾液的明显蛋白水解活性。这种活性在很大程度上是由于这种体液中明显丧失完整蛋白的损失。富含脯氨酸的蛋白质（PRP），Statherin和组织素的特征在于与矿物质稳态和抗菌活性有关的特性。尽管PRP，Statherin和Hestatin在最相关的唾液蛋白中被认为是功能上的，但它们也属于那些在释放到口腔时非常容易受到蛋白水解的唾液蛋白。我们的假设是，从腺体排泄管道释放出来的蛋白质的结构变化，以及这些蛋白质与口腔中存在的液体和细胞成分的混合将具有重要功能，从而有生理的后果。该提案的重点是唾液蛋白水解对PRP-1，Statherin，Hestatin-1和Hestatin-3的结构和功能的影响：1。研究唾液蛋白降解的速率和模式，使用AS酶的酶源液体和整个唾液的细胞分数。降解产物将通过RPHPLC进行色谱分离，并且将通过质谱法鉴定和表征各个肽。 2。确定整个唾液蛋白水解对与口腔健康直接相关的蛋白质功能的影响。为了实现这一目标，将在评估其抑制原发性和继发磷酸钙沉淀的能力以及抗真菌和抗菌活性的能力中评估蛋白质降解混合物和个体片段。项目叙述：为了维持口腔健康，适当的唾液功能是先决条件。当前的建议将研究唾液蛋白水解酶如何影响四种突出和生物学上重要的腺唾液蛋白的结构和功能。