SEPARATION OF HISTONES & CHARC OF INTACT PROTEINS &MODIFICATIONS BY MASS SPEC

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 组蛋白是一组高度保守的蛋白质，在染色质核小体中被DNA紧密包裹。它们富含修饰，如乙酰化、甲基化、磷酸化、单泛素化、糖基化和ADP，这些修饰影响DNA的可及性、其结构和功能。关于组蛋白中这些翻译后修饰的知识对于理解涉及DNA的分子过程至关重要。组蛋白的各个家族中存在的各种修饰使得全局分析困难且耗时。因此，需要能够分离各种异构体的简单可靠的方法来帮助阐明它们的修饰。 该研究项目旨在开发用于分离各种类型的组蛋白和每种类型中的异构体的方法，以降低样品的复杂性。这将使用凝胶电泳、反相和HILIC色谱的组合来实现。希望这些简化的样品能够通过FT-ICR或Orbitrap MS鉴定每个组蛋白内的翻译后修饰。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Histones are a group of highly conserved proteins wrapped tightly by DNA in chromatin nucleosomes. They are rich in modifications such as acetylation, methylation, phosphorylation, mono-ubiquitylation, glycosylation, and ADP that influence the accessibility of DNA, its architecture and functions. Knowledge about these post-translational modifications in histones is vital to understanding the molecular processes involving DNA. The variety of modifications present in the various families of histones makes global analysis difficult and time consuming. Therefore, simple and reliable methods capable of their separating the various isoforms are desirable to aid in the elucidation of their modifications. This research project is directed at the development of methods for the separation of the various types of histones and the isoforms within each type to reduce sample complexity. This will be achieved using a combination of gel electrophoresis, reverse phase and HILIC chromatography. It is hoped that these simplified samples will enable the identification of post-translational modifications within each histone by FT-ICR or Orbitrap MS.