ANALYSIS OF THE PROTEOME IN HUMAN PITUITARY TUMORS

人垂体瘤蛋白质组分析

• 批准号: 6781823
• 负责人: DOMINIC M DESIDERIO
• 金额: $ 31.36万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-15 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6781823
• 关键词: adenoma; bioinformatics; computer data analysis; digital imaging; electrofocusing; electrospray ionization mass spectrometry; epitope mapping; human tissue; mass spectrometry; matrix assisted laser desorption ionization; molecular biology information system; oncoproteins; pituitary neoplasms; posttranslational modifications; protein sequence; protein structure function; proteomics; two dimensional gel electrophoresis; western blottings

DESCRIPTION: (Adapted from applicant's abstract) The human pituitary proteome includes all proteins that are expressed in the given time. Unlike a genome, the proteome is not fixed, but it changes depending on the conditions. Proteome science (proteomics) employs a combination of state-of-the-art separation techniques, mass spectrometry, and bioinformatics for the study of proteomes. A promising prospect is the application of proteome research for the investigation of tumor pathologies. Qualitative and quantitative changes in the proteome that are associated with tumor formation can be monitored by a comparison of healthy (control) vs. tumor tissue, and the proteins of interest can be characterized, Here, the proteomics approach will be applied to the study of the proteome in human pituitary macroadenomas (hormonally silent tumors). The objective of the proposed research program is to generate a two-dimensional reference map of human pituitary proteins, and to characterize the proteins whose expression is modified in pituitary macroadenomas. The proteins will be separated by two-dimensional gel electrophoresis, and the 2D maps will be digitized. 2D gel images from control versus tumor proteomes will be compared, and protein spots will be quantified. The proteins whose abundance is altered in pituitary tumors will be identified and characterized. The protein of interest will be digested, and the masses of the resulting peptides will be measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Amino acid sequence of the peptides will be determined by electrospray-ion trap mass spectrometry. Using these peptide-mapping and/or sequencing data, the protein will be identified by searching a protein sequence database. Characterization of differentially expressed pituitary proteins will also include a detailed investigation of changes in posttranslational protein modifications. In addition, a number of prominent proteins from the control pituitary proteome will also be characterized, and a 2D reference database of the human pituitary will be constructed. Knowledge of the changes in protein expression in pituitary macroadenomas will help clarify fundamental mechanisms of macroadenoma formation, which may lead to the development of more effective means to treat and manage these tumors.

描述：（改编自申请人摘要）人垂体蛋白质组 包括在给定时间内表达的所有蛋白质。与基因组不同， 蛋白质组不是固定的，而是根据条件而变化。蛋白组 科学（蛋白质组学）采用了最先进的分离技术， 用于蛋白质组研究的技术、质谱法和生物信息学。一 蛋白质组学研究的应用是一个很有前途的前景。 肿瘤病理学研究。质量和数量的变化 与肿瘤形成相关的蛋白质组可以通过 健康（对照）与肿瘤组织的比较，以及目标蛋白质 可以表征，在这里，蛋白质组学方法将被应用于 垂体大腺瘤（垂体无瘤）蛋白质组的研究 肿瘤）。拟议研究计划的目标是产生一个 人垂体蛋白质的二维参考图谱，并表征 垂体大腺瘤中表达改变的蛋白质。 蛋白质将通过二维凝胶电泳分离， 2D地图将被数字化。对照与肿瘤蛋白质组的2D凝胶图像 将进行比较，并对蛋白质点进行定量。这些蛋白质 在垂体肿瘤中丰度改变将被识别和表征。 感兴趣的蛋白质将被消化，产生的蛋白质的质量将被消化。 肽将通过基质辅助激光解吸/电离来测量 飞行时间质谱法肽的氨基酸序列将是 通过电喷雾-离子阱质谱法测定。使用这些 肽图谱和/或测序数据，蛋白质将通过 搜索蛋白质序列数据库。差异表征 表达的垂体蛋白还将包括详细的研究， 翻译后蛋白质修饰的变化。此外，一些 来自对照垂体蛋白质组的突出蛋白质也将被 特征，并将人类垂体的2D参考数据库 构建了 了解垂体大腺瘤中蛋白质表达的变化将有助于 有助于阐明大腺瘤形成的基本机制， 发展更有效的手段来治疗和管理这些肿瘤。