Identification of nuclear type II [(3)H]estradiol binding sites as histone H4.
Identification of nuclear type II [(3)H]estradiol binding sites as histone H4.
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核 II 型 [(3)H]雌二醇结合位点鉴定为组蛋白 H4。
DOI：
10.1016/s0006-291x(02)02042-9
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发表时间：
2002
影响因子：
3.1
通讯作者：
Markaverich,BarryM
中科院分区：
文献类型：
--
作者：
Shoulars,Kevin;Brown,Trellis;Alejandro,MaryAnn;Crowley,Jan;Markaverich,BarryM
[3 H]木犀草素与子宫核II型位点共价结合[B. Markaverich，K. Shoulars，文学硕士Alejandro，T. Brown，Steroids 66（2001）707]，并用于鉴定该蛋白。[3 H]木犀草素标记的II型网站制剂的SDS-PAGE分析显示，11-和35-kDa的蛋白质的特异性结合。经氨基酸序列测定，该蛋白分子量为11 kDa，为组蛋白H4。Western blotting证实，11-和35-kDa的蛋白质是乙酰化形式的组蛋白H4。抗组蛋白H4抗体（但不是H2 A、H2 B或H3抗体）定量免疫吸附来自核提取物的II型结合位点。结合分析[3 H]雌二醇交换，使用毛地黄黄酮作为竞争对手，检测到特定的II型结合活性组蛋白H4（但不是组蛋白H2 A，H2 B，或H3）中产生的兔网织红细胞裂解物翻译系统，并确认组蛋白H4是II型网站。
[3H]Luteolin binds covalently to uterine nuclear type II sites [B. Markaverich, K. Shoulars, M.A. Alejandro, T. Brown, Steroids 66 (2001) 707] and was used to identify this protein(s). SDS–PAGE analyses of [3H]luteolin-labeled type II site preparations revealed specific binding to 11- and 35-kDa proteins. The 11-kDa protein was identified as histone H4 by amino acid sequencing. Western blotting confirmed that the 11- and 35-kDa proteins were acetylated forms of histone H4. Anti-histone H4 antibodies (but not H2A, H2B, or H3 antibodies) quantitatively immunoadsorbed type II binding sites from nuclear extracts. Binding analyses by [3H]estradiol exchange, using luteolin as a competitor, detected specific type II binding activity to histone H4 (but not histones H2A, H2B, or H3) generated in a rabbit reticulocyte lysate translation system and confirmed that histone H4 is the type II site.