Immunoproteomics

免疫蛋白质组学

• 批准号: 6772583
• 负责人: MICHAEL D. BOYLE
• 金额: $ 18.53万
• 依托单位: JUNIATA COLLEGE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6772583
• 关键词: Streptococcus infection; Streptococcus pyogenes; antigen antibody reaction; apoptosis; bacterial proteins; cell line; cysteine endopeptidases; enzyme linked immunosorbent assay; gene expression; interleukin 1; mass spectrometry; method development; molecular weight; permease; plasminogen activator; posttranslational modifications; proteomics; streptokinase; virulence; zymogens

DESCRIPTION (provided by applicant): The proposed studies are designed to evaluate new methods using a mass spectral readout to provide sensitive, detection of selected proteins, as well as methods to monitor post-translational modification events of targeted antigens. The proposed assay involves an antigen capture step mediated by immobilized antibody (immuno) and an analytical step involving mass spectral analysis of bound antigen (proteomics). The goal of the project is to develop rapid sensitive methods of antigen capture from complex mixtures of unrelated proteins in a maimer that permits the subsequent precise molecular weight determination of the bound antigen using time of flight mass spectrometry. The ability to distinguish subtle variation in the size of a targeted antigen will allow analysis of post-translational modification events for any targeted antigen to be achieved. In addition, the ability to obtain semi-quantitative data based on the area under a specific molecular weight peak on the mass spectral read-out will be critically evaluated. The proof of concept studies will focus on a number of properties of the secreted streptococcal cysteine protease SpeB which is known to post-translationally modify the surface anti-phagocytic M protein, and degrade the secreted bacterial plasminogen activator SK.

描述（由申请人提供）：拟议的研究旨在评估使用质谱读数的新方法，以提供对选定蛋白质的敏感检测，以及监测靶向抗原翻译后修饰事件的方法。提议的检测包括一个由固定抗体（免疫）介导的抗原捕获步骤和一个涉及结合抗原（蛋白质组学）的质谱分析的分析步骤。该项目的目标是开发快速灵敏的抗原捕获方法，从不相关蛋白质的复杂混合物中捕获抗原，允许随后使用飞行时间质谱法精确测定结合抗原的分子量。区分靶抗原大小的细微变化的能力将允许对任何靶抗原的翻译后修饰事件进行分析。此外，根据质谱读出的特定分子量峰下面积获得半定量数据的能力将被严格评估。概念验证研究将集中在分泌的链球菌半胱氨酸蛋白酶SpeB的一些特性上，已知SpeB在翻译后修饰表面抗吞噬M蛋白，并降解分泌的细菌纤维溶酶原激活剂SK。

项目成果

Immunoglobulin cleavage by the streptococcal cysteine protease IdeS can be detected using protein G capture and mass spectrometry.

可以使用蛋白 G 捕获和质谱法检测链球菌半胱氨酸蛋白酶 IdeS 对免疫球蛋白的裂解。

• DOI: 10.1016/j.mimet.2007.04.017
• 发表时间: 2007
• 期刊: Journal of microbiological methods
• 影响因子: 2.2
• 作者: Hess,JenniferL;Porsch,EricA;Shertz,CeceliaA;Boyle,MichaelDP
• 通讯作者: Boyle,MichaelDP

Use of protein chip mass spectrometry to monitor biotinylation reactions.

使用蛋白质芯片质谱法监测生物素化反应。

• DOI: 10.1007/s00253-006-0710-1
• 发表时间: 2007
• 期刊: Applied microbiology and biotechnology
• 影响因子: 5
• 作者: Blazer,LeviL;Boyle,MichaelDP
• 通讯作者: Boyle,MichaelDP

Practical applications of high-affinity, albumin-binding proteins from a group G streptococcal isolate.

来自 G 组链球菌分离株的高亲和力白蛋白结合蛋白的实际应用。

• DOI: 10.1007/s00253-005-0097-4
• 发表时间: 2006
• 期刊: Applied microbiology and biotechnology.
• 作者: Coyle,EmilyM;Blazer,LeviL;White,AbbyA;Hess,JenniferL;Boyle,MichaelDP
• 通讯作者: Boyle,MichaelDP