ID OF RAT UTERINE NUCLEAR TYPE II [3H]ESTRADIOL BINDING SITES AS HISTONE H4

组蛋白 H4 型大鼠子宫核 II 型 [3H]雌二醇结合位点的鉴定

• 批准号: 7355181
• 负责人: Kevin W Shoulars
• 金额: $ 0.59万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7355181
• 关键词: ID; RAT; UTERINE; NUCLEAR; TYPE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. [3H]luteolin binds nuclear type II [3H]estradiol binding sites covalently (Steroids 66707-719, 2001) and was used to identify the protein(s). Analyses of [3H]luteolin-labeled type II sites on SDS PAGE demonstrated specific binding to 11-kDa and 35-kDa proteins that were also detected in type II site preparations purified by C18-reversed phase HPLC. The 11-kDa protein was sequenced and identified as histone H4 and Western blotting confirmed the 11-kDa and 35-kDa proteins were acetylated forms of histone H4. Anti-histone H4 antibodies (but not H2A, H2B or H3 antibodies) immunoadsorbed type II sites from nuclear extracts and this was neutralized by specific competing peptides. Rabbit reticulocyte lysate translated histone H4, but not histones H2A, H2B or H3, demonstrated specific [3H]estradiol binding. Thus, histone H4 is the type II site and regulatory ligands such as methyl p-hydroxyphenyllactate (MeHPLA) and luteolin may control cell proliferation via modulation of hi stone H4 function.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。[3 H]毛地黄黄酮共价结合核II型[3 H]雌二醇结合位点（Steroids 66707-719，2001），并用于鉴定蛋白质。[3 H]木犀草素标记的SDS PAGE II型网站的分析表明，11-kDa和35-kDa的蛋白质，也检测到在II型网站的制备纯化C18-反相HPLC的特异性结合。对11-kDa蛋白进行测序并鉴定为组蛋白H4，Western印迹证实11-kDa和35-kDa蛋白是组蛋白H4的乙酰化形式。抗组蛋白H4抗体（而不是H2 A，H2 B或H3抗体）免疫吸附的II型网站从核提取物，这是由特定的竞争肽中和。兔网织红细胞裂解物翻译的组蛋白H4，而不是组蛋白H2 A，H2 B或H3，表现出特异性[3 H]雌二醇结合。因此，组蛋白H4是II型位点，调节配体（例如对羟基苯乳酸甲酯（MeHPLA）和毛地黄黄酮）可以通过调节组蛋白H4功能来控制细胞增殖。