Proteomic analysis of laser capture microscopy purified myotendinous junction regions from muscle sections
Proteomic analysis of laser capture microscopy purified myotendinous junction regions from muscle sections
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DOI：
10.1186/1477-5956-12-25
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发表时间：
2014-05-07
期刊：
影响因子：
2
通讯作者：
Blanco, Gonzalo
中科院分区：
文献类型：
--
作者：
Can, Tugba;Faas, Laura;Blanco, Gonzalo
肌腱连接是肌纤维的一种特殊结构，富含机械感应复合物，包括costameric蛋白和z盘的核心元件。在这里，激光捕获显微切割应用于纯化膜区域从小鼠骨骼肌的肌腱接头，然后进行蛋白质组学分析。肌纤维纵轴的肌膜切片用作连接制备特异性的对照。组合列表的基因本体论术语分析表明膜相关蛋白的统计学显著富集。肌腱连接制备物含有以前未表征的蛋白质、许多z盘肋配体（例如，辅肌动蛋白、capZ、aB cristallin、细丝蛋白C、cypher、钙肌蛋白、结蛋白、FHL 1、telethonin、星云蛋白、肌联蛋白和一种谜样蛋白）和其他提出的肌节拉伸传感和信号传导的参与者，如肌球蛋白和三种肌节蛋白同源物。通过免疫荧光分析证实了一个子集在肌腱连接处富集，这表明从肌肉切片中进行激光捕获显微切割是识别可能参与机械转导途径的新型肌腱连接参与者的有效方法。
The myotendinous junction is a specialized structure of the muscle fibre enriched in mechanosensing complexes, including costameric proteins and core elements of the z-disc. Here, laser capture microdissection was applied to purify membrane regions from the myotendinous junctions of mouse skeletal muscles, which were then processed for proteomic analysis. Sarcolemma sections from the longitudinal axis of the muscle fibre were used as control for the specificity of the junctional preparation. Gene ontology term analysis of the combined lists indicated a statistically significant enrichment in membrane-associated proteins. The myotendinous junction preparation contained previously uncharacterized proteins, a number of z-disc costameric ligands (e.g., actinins, capZ, aB cristallin, filamin C, cypher, calsarcin, desmin, FHL1, telethonin, nebulin, titin and an enigma-like protein) and other proposed players of sarcomeric stretch sensing and signalling, such as myotilin and the three myomesin homologs. A subset were confirmed by immunofluorescence analysis as enriched at the myotendinous junction, suggesting that laser capture microdissection from muscle sections is a valid approach to identify novel myotendinous junction players potentially involved in mechanotransduction pathways.