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Proteomic profiling of saliva for the detection of cancer.

机译：唾液的蛋白质组学分析，用于检测癌症。

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摘要

Whole saliva (WS) is the fluid that bathes the oral cavity. There has been growing interest in applying proteomics and mass spectrometry tools to body fluids in search of biomarkers for human diseases. Saliva is an attractive diagnostic fluid for such studies because it is readily available, can be collected non-invasively, and is easily stored and processed. For a meaningful comparison of the saliva in a diseased and control (healthy) state, it is essential to build a reference catalog of salivary proteins and their posttranslational modifications. The focus of my doctoral work is to catalog the salivary proteins, map the salivary glycoproteome and utilize saliva to discover potential biomarkers for pancreatic cancer.;My efforts on mapping the salivary proteome made use of two-dimensional gel electrophoresis to separate salivary proteins and mass spectrometry to identify them. We visualized more than 300 spots corresponding to salivary proteins on a 2D-gel. Of these, 105 prominent spots were excised, proteins therein digested and identified by mass spectrometry. Using this methodology, 64 proteins were discovered in whole saliva.;We mapped the salivary N-linked glycoproteome using a hydrazide based purification methodology. N-linked glycoproteins were isolated by two different methods. In the first method, glycoproteins were affixed onto a hydrazide agarose solid support. Glycoproteins were subsequently digested in situ. In the second method glycoproteins were digested before glycopeptides were affixed onto hydrazide agarose. Formerly N-linked glycopeptides were then released with the enzyme PNGase F and analyzed by mass spectrometry. The number of formerly N-glycosylated peptides and N-glycoproteins identified in WS, PA, SM, and SL fluids are 215/65 (peptides/proteins), 96/38, 114/48, and 158/54 respectively. A total of 278 formerly N-glycosylated peptides from 82 salivary N-glycoproteins have been identified.;We also applied the glycoprotein isolation method to look for protein markers for pancreatic cancer in whole saliva. Whole saliva from control subjects, patients with pancreatic cancer or pancreatitis were collected. We observed 6 N-glycoproteins that were unique to saliva from pancreatic cancer patients. We demonstrate that saliva potentially can be used as a diagnostic fluid to search for putative markers for systemic diseases as well.
机译：全唾液（WS）是沐浴口腔的液体。人们对将蛋白质组学和质谱分析工具应用于体液以寻找人类疾病的生物标记物的兴趣日益浓厚。唾液对于此类研究是有吸引力的诊断液，因为唾液易于获得，可以以非侵入方式收集，并且易于存储和处理。为了对处于患病状态和处于控制状态（健康）的唾液进行有意义的比较，必须建立唾液蛋白及其翻译后修饰的参考目录。我的博士工作重点是对唾液蛋白进行分类，绘制唾液糖蛋白图谱并利用唾液发现胰腺癌的潜在生物标志物。光谱法鉴定它们。我们在2D凝胶上可视化了300多个与唾液蛋白相对应的斑点。其中，切除了105个显着的斑点，其中的蛋白质被消化并通过质谱鉴定。使用这种方法，在整个唾液中发现了64种蛋白质。我们使用基于酰肼的纯化方法绘制了唾液N-连接的糖蛋白组图。通过两种不同的方法分离N-连接的糖蛋白。在第一种方法中，将糖蛋白固定在酰肼琼脂糖固体支持物上。糖蛋白随后被原位消化。在第二种方法中，先将糖蛋白消化，然后再将糖肽固定在酰肼琼脂糖上。然后，以前的N-连接糖肽与PNGase F酶一起释放，并通过质谱分析。在WS，PA，SM和SL液体中鉴定出的先前N-糖基化的肽和N-糖蛋白的数量分别为215/65（肽/蛋白），96 / 38、114 / 48和158/54。从82个唾液N-糖蛋白中总共鉴定出278个以前的N-糖基化肽。我们还应用糖蛋白分离方法在整个唾液中寻找胰腺癌的蛋白标记。收集来自对照受试者，胰腺癌或胰腺炎患者的全唾液。我们观察到了6种N-糖蛋白，它们是胰腺癌患者唾液特有的。我们证明，唾液潜在地可以用作诊断流体，以寻找系统性疾病的公认标志物。

著录项

  • 作者

    Ramachandran, Prasanna.;

  • 作者单位

    University of California, Los Angeles.;

  • 授予单位 University of California, Los Angeles.;
  • 学科 Chemistry Biochemistry.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 261 p.
  • 总页数 261
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

  • 入库时间 2022-08-17 11:38:30

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