Proteomic analysis of pharmacological preconditioning -: Novel protein targets converge to mitochondrial metabolism pathways
Proteomic analysis of pharmacological preconditioning -: Novel protein targets converge to mitochondrial metabolism pathways
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DOI：
10.1161/01.res.0000243995.74395.f8
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发表时间：
2006-09-29
影响因子：
20.1
通讯作者：
Van Eyk, Jennifer E.
中科院分区：
文献类型：
--
作者：
Arrell, D. Kent;Elliott, Steven T.;Van Eyk, Jennifer E.
缺血预处理的特点是对先前短暂缺血发作的缺血再灌注损伤具有抵抗作用，这种保护作用可以在体外模拟。潜在的保护机制仍然存在争议，需要更深入的了解，才能充分利用治疗。为了研究预处理对心肌蛋白质组的总体影响，用已知诱导预处理的药物腺苷或二氮嗪（各100 μ mol/L，持续60分钟）处理离体兔心室肌细胞。然后使用多层2D凝胶电泳方法将其蛋白质谱与溶剂处理对照组（每次处理n = 4只动物）进行比较。在28个显著改变的蛋白质点中，鉴定了19个非冗余蛋白质（5个点仍未鉴定）。这些蛋白质中的大多数参与线粒体能量学，包括三羧酸循环酶和氧化磷酸化复合物的亚基。这些变化不是不加区别的，只有少量的酶或复杂的亚基改变，表明这2个预处理模拟物的非常具体和有针对性的影响。在这些变化中，有4种蛋白质的翻译后修饰程度发生了变化。其中之一，腺苷诱导的磷酸化的ATP合成酶β亚基，充分其特征在于与5个新的磷酸化位点的鉴定。这种蛋白质组学方法提供了一个整体评估的细胞反应与腺苷和二氮嗪的药理学治疗，并确定了一个独特的子集的酶和蛋白质复合物亚基，可能是基础的预处理表型。
Ischemic preconditioning is characterized by resistance to ischemia reperfusion injury in response to previous short ischemic episodes, a protective effect that can be mimicked pharmacologically. The underlying mechanism of protection remains controversial and requires greater understanding before it can be fully exploited therapeutically. To investigate the overall effect of preconditioning on the myocardial proteome, isolated rabbit ventricular myocytes were treated with drugs known to induce preconditioning, adenosine or diazoxide ( each at 100 mu mol/L for 60 minutes). Their protein profiles were then compared with vehicle-treated controls (n = 4 animals per treatment) using a multitiered 2D gel electrophoresis approach. Of 28 significantly altered protein spots, 19 nonredundant proteins were identified ( 5 spots remained unidentified). The majority of these proteins are involved in mitochondrial energetics, including subunits of tricarboxylic acid cycle enzymes and oxidative phosphorylation complexes. These changes were not indiscriminate, with only a small number of enzymes or complex subunits altered, indicating a very specific and targeted affect of these 2 preconditioning mimetics. Among the changes were shifts in the extent of posttranslational modification of 4 proteins. One of these, the adenosine-induced phosphorylation of the ATP synthase beta subunit, was fully characterized with the identification of 5 novel phosphorylation sites. This proteomics approach provides an overall assessment of the cellular response to pharmacological treatment with adenosine and diazoxide and identifies a distinct subset of enzymes and protein complex subunit that may underlie the preconditioned phenotype.