Automated Glyco-Analysis of Cancer Related Proteins

癌症相关蛋白的自动糖分析

• 批准号: 7140158
• 负责人: Lewis K. Pannell
• 金额: $ 14.26万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-18 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7140158
• 关键词: biomarker; cancer information system; cell surface receptors; computer program /software; glycoprotein biosynthesis; glycoprotein structure; glycosylation; mass spectrometry; membrane proteins; neoplastic growth; posttranslational modifications; proteomics; technology /technique development

DESCRIPTION (provided by applicant): The quote that "aberrant glycosylation is the hallmark of cancer cells" is reflected in numerous reports in the literature documenting changes in glycosylation on specific membrane proteins in cancer cells relative to normal cells. These changes have been shown to be involved in the release of cancer cells into the extracellular matrix and in the formation of metastasis. Glycosylated proteins represent a huge, almost untapped source of biomarkers, considering the wealth of evidence documenting their significance in cancer. Unique glycoforms could be used for diagnostic purposes, to target drugs at cancer cells, and for the development of immunotherapy. Despite the evolution of new mass spectrometry based methods for protein analysis, few of these involve the determination of post-translational modifications, especially glycosylation. As routine methods (e.g., MS/MS based sequencing methods) yield little light on glycosylated peptides, this proteomics research facility has established a new approach to automatically identify glycan structures on pure proteins from commercial or recombinant sources. It involves the acquisition of molecular weight only spectra and the detection of the glycosylation patterns using accurately determined mass gaps between the various glycoforms. The presence of multiple glycoforms is used to enhance the analysis rather than to confuse it. The approach has been shown to be reliable and extremely fast (taking less than one second) at identifying and characterizing such sites, including in proteins with highly complex glycosylation patterns. The aim of this proposal is to prove its utility in cancer where changes in glycosylation changes are interlaced with the progression of the disease. It will concentrate on both the cell surface proteins and those secreted from cells. Data will be compared to previously published reports where available. The glycans on previously uncharacterized proteins will be established and validated against the best hand interpreted results. The long-term aim is to make glycoanalyses routine to all cancer investigators, and the software integral to the approach will be made publicly available on a www site. This will represent the first step, with the glyco-analysis of full proteomes being an ultimate objective.

描述（由申请人提供）：“异常糖基化是癌细胞的标志”这句话反映在大量文献报道中，这些文献记录了癌细胞中特定膜蛋白糖基化相对于正常细胞的变化。这些变化已被证明与癌细胞释放到细胞外基质和转移的形成有关。考虑到大量证据证明糖基化蛋白在癌症中的重要性，糖基化蛋白代表了一个巨大的、几乎未开发的生物标志物来源。独特的糖型可用于诊断目的，靶向药物针对癌细胞，并为免疫治疗的发展。尽管新的基于质谱的蛋白质分析方法不断发展，但这些方法很少涉及到翻译后修饰的测定，特别是糖基化。由于常规方法（例如，基于质谱/质谱的测序方法）对糖基化肽的研究很少，该蛋白质组学研究设施已经建立了一种自动识别来自商业或重组来源的纯蛋白质上的聚糖结构的新方法。它包括仅获得分子量光谱和使用精确确定的各种糖型之间的质量间隙来检测糖基化模式。多种糖型的存在是用来加强分析，而不是混淆它。该方法已被证明是可靠的，并且在识别和表征这些位点（包括具有高度复杂的糖基化模式的蛋白质）方面非常快速（不到一秒钟）。这项提议的目的是证明其在癌症中的效用，其中糖基化变化的变化与疾病的进展交织在一起。它将集中于细胞表面蛋白质和细胞分泌的蛋白质。如有数据，将与以前发表的报告进行比较。先前未表征的蛋白质上的聚糖将根据最佳的手解释结果建立和验证。长期目标是使糖分析成为所有癌症研究人员的日常工作，而与该方法相结合的软件将在www网站上公开。这将是第一步，完整蛋白质组的糖分析是最终目标。

项目成果

Large isoform of MRJ (DNAJB6) reduces malignant activity of breast cancer.

• DOI: 10.1186/bcr1874
• 发表时间: 2008
• 期刊: BREAST CANCER RESEARCH
• 影响因子: 7.4
• 作者: Mitra, Aparna;Fillmore, Rebecca A.;Metge, Brandon J.;Rajesh, Mathur;Xi, Yaguang;King, Judy;Ju, Jingfang;Pannell, Lewis;Shevde, Lalita A.;Samant, Rajeev S.
• 通讯作者: Samant, Rajeev S.