PROTEINS IN THE ORAL FLUIDS AND TOOTH PELLICLE LAYER

口腔液和牙膜层中的蛋白质

• 批准号: 7722973
• 负责人: Frank G Oppenheim
• 金额: $ 0.07万
• 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: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722973
• 关键词: Adsorption; Amylases; Antibodies; Blood capillaries; Chromatography; Computer Retrieval of Information on Scientific Projects Database; Consult; Consultations; Coupled; Cystatins; Data; Databases; Degradation Pathway; Dental Enamel; Dental Pellicle; Depth; Detection; Film; Funding; Gel; Gingiva; Grant; Human; Human Resources; Hydroxyapatites; Institution; Ion-Exchange Chromatography Procedure; Keratin; Lead; Leukocyte L1 Antigen Complex; Liquid substance; Methods; Operative Surgical Procedures; Oral; Oral cavity; Peptides; Post-Translational Protein Processing; Property; Protein Analysis; Protein Biosynthesis; Proteins; Publishing; Radiolabeled; Research; Research Personnel; Resources; Saliva; Salivary; Salivary Proteins; Serum; Source; Spottings; System; Time; Tooth structure; United States National Institutes of Health; capillary; cystatin SN; data acquisition; gel electrophoresis; in vivo; liquid chromatography mass spectrometry; proline-rich proteins; radiotracer; saliva secretion; statherin; tooth surface

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The acquired enamel pellicle is the organic film primarily derived from salivary components that covers and protects the enamel surface of teeth. The ultimate aim of this project is to characterize the composition and functional properties of the acquired enamel pellicle. Previous studies, using methods including gel electrophoresis, classical protein separation chromatography, and antibody detection were limited by the small amounts of material. These methods suggested that the pellicle is composed of one component or another in amounts difficult to relate to the pellicle as a whole. Past efforts, in particular adsorption studies using radiolabelled proteins and mass spectral analysis of pellicle proteins separated by 2-D PAGE, provided information indicating a high content of phosphorylated proteins. Additionally, proline-rich proteins, statherin and cystatin SN, were identified in artificial pellicles produced by adsorption of salivary secretions to hydroxyapatite. However, when a similar analysis of proteins derived from in vivo collected pellicle was conducted, the proline-rich proteins were notably absent. Characterization of 2D gel spots from saliva and in vivo pellicle was performed by analyzing extracted tryptic peptides. Database searches using both MS and MS/MS data obtained using MALDI-TOF and Q-oTOF MS identified expected salivary proteins (cystatins, amylase, statherin, and others), as well as unanticipated gingival serum fluid proteins (i.e., calgranulin) and cellular proteins (endothelial keratins). Our results from this study were published in J. Biol. Chem. In order to provide an even more comprehensive picture of salivary and pellicle components, ion exchange chromatography coupled with on-line capillary LC/MS has been employed. To facilitate this analysis, a defined database containing all the known oral proteins and other components that can be present in the oral cavity has been constructed and the resuls from the salivary proeome study are being consulted. Data acquired during intelligent data acquisition (IDA) operation of the capLC-QStar i nanoESI-orthogonal TOF MS system can be quickly searched against the available databases, and searches that allow for all known post-translational modifications can be performed rapidly and reliably. Use of this approach has significantly increased the assignments of eluting peaks and has yielded highly reliable and consistently relevant results. This approach should lead to an in-depth understanding of protein synthesis/degradation pathways and functions in oral fluids and human pellicle. Prof. Oppenheim's department recently acquired its own LTQ MS in order to dedicate more time to these analyses. Resource personnel will continue to provide advice and consultation.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 获得的搪瓷膜是主要源自唾液成分的有机膜，该成分覆盖和保护牙齿的牙釉质表面。 该项目的最终目的是表征所获得的搪瓷颗粒的组成和功能特性。 先前的研究，使用包括凝胶电泳，经典蛋白分离色谱和抗体检测的方法受到少量材料的限制。这些方法表明，颗粒由一个或另一个组成的量组成，量很难与整个颗粒相关。 过去的努力，特别是使用放射性标记蛋白的吸附研究和对二-D页分离的细胞蛋白的质谱分析，提供了表明磷酸化蛋白质含量高的信息。 另外，在通过吸附唾液中分泌到羟基磷灰石产生的人造脂肪中鉴定出富含脯氨酸的蛋白，Statherin和胱抑素SN。但是，当对从体内收集的颗粒中衍生的蛋白质进行类似的分析时，明显没有脯氨酸的蛋白质。 通过分析提取的胰蛋白酶肽进行了来自唾液和体内颗粒的2D凝胶斑点的表征。 使用使用MALDI-TOF和Q-OTOF MS获得的MS和MS/MS数据的数据库搜索确定了预期的唾液蛋白（胱抑素，淀粉酶，Statherin等），以及意外的牙龈血清液体蛋白（即Calgranulin）（即Calgranulin）和细胞蛋白（calgranulin）和细胞蛋白（Endophartar蛋白（Endopharlar蛋白）。这项研究的结果发表在J. Biol中。化学 为了提供有关唾液和颗粒成分的更全面的图片，已经采用了离子交换色谱以及在线毛细管LC/MS的情况。为了促进这种分析，已经构建了一个包含所有可存在于口腔中的已知口服蛋白和其他成分的数据库，并咨询了唾液proeme研究中的重新验证。可以在可用数据库中快速搜索CAPLC-QSTAR I正交TOF MS系统的智能数据采集（IDA）操作中获取的数据，并且可以快速且可靠地进行所有已知的翻译后修改的搜索。这种方法的使用显着增加了洗脱峰的分配，并产生了高度可靠且一致相关的结果。 这种方法应导致对蛋白质合成/降解途径的深入理解，并在口服流体和人膜中的功能。 Oppenheim教授最近获得了自己的LTQ MS，以便将更多时间用于这些分析。资源人员将继续提供建议和咨询。