| 张勤 薛鹏 白宝玲 李会莉 杨福全.质谱检测叶酸缺乏人胚肾细胞中低组蛋白H3赖氨酸79甲基化修饰[J].,2014,14(23):4420-4424 |
| 质谱检测叶酸缺乏人胚肾细胞中低组蛋白H3赖氨酸79甲基化修饰 |
| Low Histone H3 Lysine 79 Methylation in Folic AcidDeficiency HEK-293 Cells by Mass Spectrometry |
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| DOI： |
| 中文关键词: H3K79 甲基化修饰 质谱 叶酸缺乏 人胚肾细胞 |
| 英文关键词: H3K79 methylation HPLC-LTQ/Orbitrap Ms FA deficiency HEK-293 |
| 基金项目:国家自然科学基金项目（81300489） |
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| 中文摘要: |
| 目的：探讨叶酸（Folic acid，FA）缺乏在培养的人胚肾细胞（HEK-293）中对细胞组蛋白修饰水平的影响。方法：人胚肾细胞分
两组培养，一组正常培养，一组无叶酸培养。细胞提取组蛋白后通过高效液相色谱一线性离子阱／静电场轨道阱高分辨质谱
（HPLC-LTQ/Orbitrap Ms）检测组蛋白的修饰以比较叶酸缺乏对人胚肾细胞组蛋白修饰的影响。结果：用高分辨质谱方法成功检测
到人胚肾细胞的五个组蛋白变体H1，H3，H4，H2a 和H2b 上的33个组蛋白修饰位点，其中23个修饰位点为uniprot 数据库上已
经报道的组蛋白修饰位点，而其余10 个为未报道修饰位点。通过质谱比较正常和叶酸缺乏组人胚肾细胞修饰谱发现H3K79me1
和H3K79me2在叶酸缺乏培养组中检出率较低。进一步用蛋白免疫印迹的方法也证明了在叶酸缺乏的人胚肾细胞中H3K79me1
水平低于正常培养组。结论：细胞中叶酸缺乏影响组蛋白甲基化包括H3K79me2 和H3K79me1 修饰水平，提示细胞外营养因素叶
酸水平可影响组蛋白修饰水平从而参与疾病如神经管畸形（Neural tube defect，NTD）的发生。 |
| 英文摘要: |
| Objective:To explore the roles of Folic acid (FA) in cultured human embryonic kidney cells (HEK-293) on histone
modification levels.Methods:The cultured human embryonic kidney cells were divided into two groups. One group was cultured with
normal DMEM, while the other group was cultured with Folate-free culture medium. Histone modification level was detected and
compared by HPLC linear ion trap/Orbitrap high-resolution mass spectrometry (HPLC-LTQ/Orbitrap Ms) between two groups.Results:33 Modification sites were found in the H1, H3, H4, H2a and H2b variants by HPLC-LTQ/Orbitrap Ms in the human embryonic kidney
cells. And the level of H3K79me1 and H3K79me2 were found significantly lower in FA deficiency group than in the control group. And
the western-blot confirmed that the H3K79me1 level was lower in Folate-free group.Conclusion:The FA deficiency in cell environment
affects histone methylation modifications which suggests neural tube defects (NTDs) may be partly due to lack of nutrition factors such as
FA. |
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