Department of Medicine, Division of Molecular Cardiology, Texas A&M Health Science Center, Temple, TX 76504, USA.
Microsc Microanal. 2012 Feb;18(1):107-14. doi: 10.1017/S1431927611012359. Epub 2011 Dec 12.
Normal cardiac function is maintained through dynamic interactions of cardiac cells with each other and with the extracellular matrix. These interactions are important for remodeling during cardiac growth and pathophysiological conditions. However, the precise mechanisms of these interactions remain unclear. In this study we examined the importance of desmoplakin (DSP) in cardiac cell-cell interactions. Cell-cell communication in the heart requires the formation and preservation of cell contacts by cell adhesion junctions called desmosome-like structures. A major protein component of this complex is DSP, which plays a role in linking the cytoskeletal network to the plasma membrane. Our laboratory previously generated a polyclonal antibody (1611) against the detergent soluble fraction of cardiac fibroblast plasma membrane. In attempting to define which proteins 1611 recognizes, we performed two-dimensional electrophoresis and identified DSP as one of the major proteins recognized by 1611. Immunoprecipitation studies demonstrated that 1611 was able to directly pulldown DSP. We also demonstrate that 1611 and anti-DSP antibodies co-localize in whole heart sections. Finally, using a three-dimensional in vitro cell-cell interaction assay, we demonstrate that 1611 can inhibit cell-cell interactions. These data indicate that DSP is an important protein for cell-cell interactions and affects a variety of cellular functions, including cytokine secretion.
正常的心脏功能是通过心肌细胞之间以及与细胞外基质的动态相互作用来维持的。这些相互作用对于心脏生长和病理生理条件下的重塑很重要。然而,这些相互作用的确切机制尚不清楚。在这项研究中,我们研究了桥粒斑蛋白(DSP)在心肌细胞间相互作用中的重要性。心脏中的细胞间通讯需要通过称为桥粒样结构的细胞黏附连接来形成和维持细胞接触。该复合物的一个主要蛋白成分是 DSP,它在将细胞骨架网络与质膜连接中起作用。我们实验室之前生成了一种针对心肌成纤维细胞质膜去垢剂可溶性部分的多克隆抗体(1611)。在试图确定 1611 识别哪些蛋白时,我们进行了二维电泳,发现 DSP 是 1611 识别的主要蛋白之一。免疫沉淀研究表明,1611 能够直接下拉 DSP。我们还证明 1611 和抗-DSP 抗体在整个心脏切片中共同定位。最后,使用三维细胞间相互作用测定,我们证明 1611 可以抑制细胞间相互作用。这些数据表明,DSP 是细胞间相互作用的重要蛋白,影响多种细胞功能,包括细胞因子分泌。