Identification of phosphoproteins in kidney tissues from patients with autosomal dominant polycystic kidney disease
Identification of phosphoproteins in kidney tissues from patients with autosomal dominant polycystic kidney disease
复制标题
常染色体显性多囊肾病患者肾组织中磷蛋白的鉴定
DOI：
10.1002/prca.200780172
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发表时间：
2008-07-01
影响因子：
2
通讯作者：
Sandford, Richard
中科院分区：
文献类型：
--
作者：
Liu, Yawei;Dai, Bing;Sandford, Richard
蛋白质磷酸化是一个非常重要的PTM。蛋白质的磷酸化/去磷酸化可以以几乎所有可能的方式改变其行为。以往的研究表明，异常磷酸化参与了常染色体显性多囊肾病（ADPKD）的发病机制。然而，尚未报道ADPKD中改变的磷蛋白的大规模蛋白质组学分析。本研究提取了ADPKD囊肿肾组织（n = 5）和正常肾组织（n = 5）的总蛋白，经磷酸金属亲和层析富集磷蛋白，然后进行双向电泳分离和LC-MS/MS鉴定。 其中，28个点在ADPKD肾组织中表达上调，20个点表达下调。其中，38个不同的蛋白质被鉴定，包括细胞信号蛋白，细胞骨架蛋白，线粒体代谢酶，抗氧化蛋白，分子伴侣，转录因子和调节因子。两个差异磷蛋白，膜联蛋白II和原肌球蛋白，进一步证实了免疫沉淀和Western印迹分析。结果表明，多囊肾病囊肿组织中存在多种异常磷酸化蛋白。进一步研究这些差异磷蛋白的功能可能为ADPKD的发病机制和治疗提供新的线索。
Protein phosphorylation is a very important PTM. Phosphorylation/dephosphorylation of a protein can alter its behavior in almost every conceivable way. Previous studies indicate that abnormal phosphorylation is involved in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). However, large‐scale proteomic analysis of altered phosphoproteins in ADPKD has not been reported. In this study, total proteins from ADPKD cystic kidney tissues (n = 5) and normal kidney tissues (n = 5) were extracted and phosphoproteins were enriched by phosphate metal affinity chromatography, then separated by 2‐DE and identified by LC‐MS/MS. Between the two groups, 48 protein spots showing more than a twofold difference were detected. Among them, 28 spots were up‐regulated and 20 down‐regulated in ADPKD kidney tissues. Of these, 38 different proteins were identified including cell signaling proteins, cytoskeleton proteins, mitochondria metabolic enzymes, antioxidant proteins, molecular chaperones, transcription factors and regulators. Two differential phosphoproteins, annexin II and tropomyosin, were further confirmed by immunoprecipitation and Western blot analysis. The results show that there are many kinds of abnormal phosphoproteins in ADPKD cystic kidney tissues. More studies on the functions of the differential phosphoproteins may provide us new clues for ADPKD pathogenesis and treatment.