| 同位素标签技术在人肾小管上皮细胞蛋白质组学研究中的应用 |
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| 引用本文： | 陈永熙,李娅,张文,陈晓农,王伟铭,张群业,黄秋花,陈楠. 同位素标签技术在人肾小管上皮细胞蛋白质组学研究中的应用[J]. 上海交通大学学报(医学版), 2009, 29(5) |
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| 作者姓名： | 陈永熙 李娅 张文 陈晓农 王伟铭 张群业 黄秋花 陈楠 |
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| 作者单位： | 陈永熙,李娅,张文,陈晓农,王伟铭,陈楠,CHEN Yong-xi,LI Ya,ZHANG Wen,CHEN Xiao-nong,WANG Wei-ming,CHEN Nan(上海交通大学,医学院瑞金医院肾脏科,上海,200025);张群业,黄秋花,ZHANG Qun-ye,HUANG Qiu-hua(上海交通大学,上海血液学研究所,医学基因组学国家重点实验室,上海,200025) |
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| 基金项目： | 国家自然科学基金，上海市重点学科项目，上海市卫生局重点课题，上海市卫生局重点学科基金 |
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| 摘 要： | 目的 建立并优化人肾小管上皮细胞蛋白质组学分析所需的相对和绝对定量的同位素标签试剂(iTRAQ)标记技术,为进一步研究奠定基础.方法 将人肾小管上皮细胞株HK-2通过细胞裂解、丙酮沉淀、蛋白变性、还原及酶解,最后进行iTRAQ标记和质谱鉴定,研究iTRAQ在HK-2细胞的标记情况和蛋白质组学.结果 通过质谱检测,检测到带有iTRAQ标记肽段.随机选取20个前体离子,通过3次重复试验,鉴定出iTRAQ标记多肽肽段分别为17、15和18个,标记效率在75.5% ～90.0%,且组间标记效率差异无统计学意义(P>0.05).结论 iTRAQ可以在HK-2细胞中标记效果满意,且重复性较好,为进一步研究肾脏疾病的蛋白质组学奠定了基础.
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| 关 键 词： | 相对和绝对定量的同位素标签试剂 定量蛋白质组学 人肾小管上皮细胞 |
Application of isobaric tags labeling in proteomic research of human renal proximal tubular epithelial cells |
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| CHEN Yong-xi,LI Ya,ZHANG Wen,CHEN Xiao-nong,WANG Wei-ming,ZHANG Qun-ye,HUANG Qiu-hua,CHEN Nan. Application of isobaric tags labeling in proteomic research of human renal proximal tubular epithelial cells[J]. Journal of Shanghai Jiaotong University:Medical Science, 2009, 29(5) |
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| Authors: | CHEN Yong-xi LI Ya ZHANG Wen CHEN Xiao-nong WANG Wei-ming ZHANG Qun-ye HUANG Qiu-hua CHEN Nan |
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| Affiliation: | 1.Department of Nephrology;2.Shanghai Institute of Hematology;National Key Laboratory of Medical Genomics;Ruijin Hospital;School of Medicine;Shanghai Jiaotong University;Shanghai 200025;China |
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| Abstract: | Objective To establish and optimize proteomic research platform using isobaric tags for relative and absolute quantitation(iTRAQ) so as to facilitate further proteomic research of human renal proximal tubular epithelial cells.Methods After cell lysis,acetone precipitation,denaturation and protein digestion of human renal proximal tubular epithelial cell line HK-2,the peptide was labeled with iTRAQ reagents and subjected to mass spectrometry.The labeling of iTRAQ in HK-2 cells and proteomics were studied. Re... |
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| Keywords: | isobaric tags for relative and absolute quantitation quantitative proteomics human renal proximal tubular epithelial cell |
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