Ser10磷酸化的组蛋白H3和微管蛋白在小麦根尖细胞有丝分裂过程中的动态分布

利用间接免疫萎光标记技术研究Ser10磷酸化的组蛋白H3和微管蛋白在小麦根尖细胞中有丝分裂过程中的动态分布情况.结果显示在小麦根尖细胞有丝分裂过程中Ser10磷酸化的组蛋白H3的出现和消失与染色体的凝集和解凝集的过程存在时空上的相关性,在有丝分裂的过程中这种蛋白在着经线粒上的定位有有助于染色体向两极移动.研究结果还表明,在有丝分裂过程中,微管蛋白发生了重组,成束的垂直排列在赤道板的两侧,协助细胞有丝分裂过程的顺利完成.     

A quantitative analysis of histone methylation and acetylation isoforms from their deuteroacetylated derivatives: application to a series of knockout mutants

The core histones, H2A, H2B, H3 and H4, undergo post‐translational modifications (PTMs) including lysine acetylation, methylation and ubiquitylation, arginine methylation and serine phosphorylation. Lysine residues may be mono‐, di‐ and trimethylated, the latter resulting in an addition of mass to the protein that differs from acetylation by only 0.03639 Da, but that can be distinguished either on high‐performance mass spectrometers with sufficient mass accuracy and mass resolution or via retention times. Here we describe the use of chemical derivatization to quantify methylated and acetylated histone isoforms by forming deuteroacetylated histone derivatives prior to tryptic digestion and bottom‐up liquid chromatography‐mass spectrometric analysis. The deuteroacetylation of unmodified or mono‐methylated lysine residues produces a chemically identical set of tryptic peptides when comparing the unmodified and modified versions of a protein, making it possible to directly quantify lysine acetylation. In this work, the deuteroacetylation technique is used to examine a single histone H3 peptide with methyl and acetyl modifications at different lysine residues and to quantify the relative abundance of each modification in different deacetylase and methylase knockout yeast strains. This application demonstrates the use of the deuteroacetylation technique to characterize modification ‘cross‐talk’ by correlating different PTMs on the same histone tail. Copyright © 2013 John Wiley & Sons, Ltd.     

组蛋白H3赖氨酸27和36位甲基化修饰的鉴定

采用液相色谱-质谱联用技术结合生物信息学分析手段,研究Hela细胞组蛋白H3赖氨酸(Lysine (K))K27和K36位点带有甲基化和二甲基化修饰的多肽鉴定,通过二级质谱碎片解析和二级碎片丰度分析,对组蛋白H3赖氨酸K27和K36上甲基化和二甲基化修饰进行了鉴定和分析。     

Force field parameters for the simulation of modified histone tails

We describe the development of force field parameters for methylated lysines and arginines, and acetylated lysine for the CHARMM all‐atom force field. We also describe a CHARMM united‐atom force field for modified sidechains suitable for use with fragment‐based docking methods. The development of these parameters is based on results of ab initio quantum mechanics calculations of model compounds with subsequent refinement and validation by molecular mechanics and molecular dynamics simulations. The united‐atom parameters are tested by fragment docking to target proteins using the MCSS procedure. The all‐atom force field is validated by molecular dynamics simulations of multiple experimental structures. In both sets of calculations, the computational predictions using the force field were compared to the corresponding experimental structures. We show that the parameters yield an accurate reproduction of experimental structures. Together with the existing CHARMM force field, these parameters will enable the general modeling of post‐translational modifications of histone tails. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2010     

新型吡啶苯磺酰胺联喹啉基异羟肟酸类PI3K/HDAC双靶点抑制剂的设计、合成和生物活性研究

多靶点药物已成为一种有广阔前景的药物,特别是对抗肿瘤药物的研发.基于候选药物GSK2126458和上市药物Vorinostat的结构特点,设计并合成了一系列新型的磷脂酰肌醇3-激酶(PI3Ks)和组蛋白脱乙酰酶(HDACs)双重抑制剂.生物活性研究发现,化合物GYB-4对PI3Kα和HDAC1的IC50分别为1.0和4.2nmol/L;化合物GYB-5对PI3Kα和HDAC1的IC50分别为1.3和4.8nmol/L.对所有化合物在HCT116,PC3和A2780细胞株上进行了增殖抑制活性研究,相关的构效关系研究将为PI3K和HDAC双靶点抑制剂的进一步优化提供思路.     

一种香豆素荧光标记多肽的合成及应用

Sirtuin蛋白是一类称为依赖烟酰胺腺嘌呤二核苷酸（NAD）的组蛋白去乙酰化酶,共有7个成员,均是潜在的疾病治疗靶点。 然而,目前的荧光筛选方法,只适用于SIRT1~SIRT3。 因此,根据SIRT5的新酶活,设计、合成了针对SIRT5的荧光标记多肽（ISGASE(SuK) AMC）,并通过LC-MS和荧光检测证明了该荧光标记多肽能应用于SIRT5的活性筛选。     

表观遗传试剂诱导滇重楼内生真菌 Hypomyces sp.CLG4次生代谢产物的研究

采用高效液相色谱(HPLC)分析添加HDACI,500μM恩提诺特的H.sp.CLG4.培养萃取物与对照的差异,硅胶柱层析、反相柱层析、半制备高效液相等色谱方法对添加表观遗传试剂的Hypomyces sp.CLG4.次级代谢产物进行分离纯化,并利用现代波谱分析方法对化合物进行结构鉴定.研究组蛋白去乙酰化酶抑制剂(his...