The C-terminome

C端组

基本信息

批准号：
8575072
负责人：
MARTIN R SCHILLER
金额：
$ 44.27万
依托单位：
UNIVERSITY OF NEVADA LAS VEGAS
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8575072
关键词：
Abbreviations Affinity Affinity Chromatography Amino Acids Automobile Driving Binding Binding Proteins Bioinformatics Biological Biological Assay Biological Process C-terminal Case Study Cataloging Catalogs Cell Extracts Cells Coin Confocal Microscopy Databases Disease Drug usage Exocytosis FDA approved Gene Family Human Internet Label Letters Mass Spectrum Analysis Mutate Mutation Names Patients Pattern Peptides Pharmaceutical Preparations Point Mutation Post-Translational Protein Processing Protein Import Protein Splicing Proteins Proteome RNA Interference Role Set protein Sucrose Synapsin II System Testing Validation Variant graduate student insight member mimetics novel peroxisome protein aminoacid sequence protein function protein transport public health relevance sedimentation velocity synaptotagmin V trafficking web page web site

项目摘要

DESCRIPTION (provided by applicant): The C-termini are often very important for protein function, often containing short contiguous peptide sequences (minimotifs) that encode post-translational modification, zipcode motifs for trafficking to specific cell compartments, and minimotifs for binding to other proteins and molecules. Through our annotation of 100,000s of minimotifs in the Minimotif Miner database our lab has catalogued >1000 known functional minimotifs specific to the carboxy (C)-termini of proteins. These C-terminal minimotifs are critica for a wide array of biological functions. Mutations can disable minimotifs by a single point mutation producing disease states and some minimotif mimetics are FDA-approved drugs used to treat patients. In this proposal we seek to identify new minimotifs and consolidate existing information into what we coin the "functional C-terminome", the general role of the C-terminus in all human proteins. We first ask the question: Are there important C-terminal minimotifs that remain to be discovered? In our preliminary studies two graduate students performed a bioinformatic analysis to identify many possible minimotif patterns that overrepresented on the C- termini of different human proteins when compared to a scrambled human proteome. We next selected four of these minimotifs and in a pilot screen an undergraduate used affinity chromatography followed by mass spectrometry to identify potential binding partners for these minimotifs. We propose to expand upon these findings by (1) determining the relevance of the PxP-carboxy terminal minimotif in stimulate exocytosis as a case study, (2) identify interactors of novel C-terminal motifs by affinity capture and mass spectrometry (LC MS/MS), and (3) to consolidate all known information about the functional C-terminome and generate a searchable webpage for exploring all C-terminal minimotifs in the human proteome.

描述（由申请人提供）：C-termini通常对于蛋白质功能非常重要，通常包含编码翻译后修饰后的短连续性肽序列（微型肽），用于运输到特定细胞室的Zipcode基序，以及与其他蛋白质和分子结合的最小值。通过我们在最小矿工数据库中对100,000秒的最小值的注释，我们的实验室已分类> 1000个已知的功能最小值，该功能最小值特定于蛋白质的羧基末端（C）末端。这些C末端的极小拟合是各种生物学功能的批评。突变可以通过产生疾病状态的单点突变来禁用最小化，而某些微型模拟物是用于治疗患者的FDA批准的药物。在此提案中，我们试图确定新的最小值，并将现有信息巩固到“功能性C末端组”中，这是C-末端在所有人类蛋白中的一般作用。我们首先提出一个问题：是否有重要的C末端极小拟合有待发现？在我们的初步研究中，两位研究生进行了生物信息学分析，以确定许多可能的极小模式，这些模式在不同人类蛋白质的c-末端过分代表了不同人类蛋白质，而与炒作的人类蛋白质组相比。接下来，我们选择了其中四个微型拟合，并在试验屏幕中，本科生使用的亲和力色谱法，然后进行质谱法，以识别这些最小值的潜在结合伴侣。 We propose to expand upon these findings by (1) determining the relevance of the PxP-carboxy terminal minimotif in stimulate exocytosis as a case study, (2) identify interactors of novel C-terminal motifs by affinity capture and mass spectrometry (LC MS/MS), and (3) to consolidate all known information about the functional C-terminome and generate a searchable webpage for exploring all C-terminal人类蛋白质组中的最小值。