Pleissner K P, Regitz-Zagrosek V, Weise C, Neuss M, Krüdewagen B, Söding P, Buchner K, Hucho F, Hildebrandt A, Fleck E
Klinik Innere Medizin, Schwerpunkt Kardiologie/Angiologie, Universitätsklinikum Rudolf Virchow, Freien Universität Berlin, Germany.
Electrophoresis. 1995 May;16(5):841-50. doi: 10.1002/elps.11501601139.
High resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), followed by computer-assisted image analysis (PDQUEST) was used to screen atrial and ventricular protein patterns for quantitative and qualitative differences in protein expression. Myocardial proteins from left ventricular (LV) and right atrial (RA) samples from end-stage, failing explanted hearts and from a healthy donor heart (control) were separated by 2-D large gel electrophoresis. Ten RA versus ten LV gels from explanted dilated cardiomyopathic (DCM) hearts were analyzed for quantitative differences in their spot patterns. Of the 197 spots matched to every gel, 40 spots differed significantly in intensity between RA and LV for DCM patients. A larger number of atrial and ventricular gels (20 RA, 20 LV) from DCM patients and from a healthy donor heart (4 RA, 4 LV gels) were analyzed for qualitative differences in protein expression. Three protein spots (SSP 1120: M(r)/pI:20.5 kDa/4.6; SSP 1119: M(r)/pI:20.6 kDa/4.5; SSP 0117:M(r)/pI:20.7/ < 4.5) that are present in all RA gels for DCM patients are absent in all LV gels. Two protein spots (SSP 0112: M(r)/pI:17.2 kDa,/ < 4.4; SSP 0114:M(r)/pI:17.6 kDa/ < 4.4) occur only in all LV gels but not in the RA gels. These five qualitatively differing spots are identical in DCM patients and in the healthy donor heart. Some of the differing spots were internally sequenced and identified as myosin light chain isoforms (myosin light chain 2, atrial; myosin light chain 2, ventricular; myosin light chain 1, atrial) with the Protein Identification Resource (PIR) accession numbers A44451, S03708, A30881, respectively. Additionally, phosphoglycerate mutase (PIR: JQ0750) and ATP synthase alpha chain (PIR: S17193) were identified. Thus, quantitative and qualitative differences between atrial and ventricular protein patterns were identified by 2-D PAGE. A characteristic distribution of myosin light chains between atrial and ventricular human myocardium was found using our approach.
采用高分辨率二维聚丙烯酰胺凝胶电泳（2-D PAGE），随后进行计算机辅助图像分析（PDQUEST），以筛选心房和心室蛋白质模式，寻找蛋白质表达在定量和定性方面的差异。通过二维大型凝胶电泳分离来自晚期衰竭心脏移植样本的左心室（LV）和右心房（RA）心肌蛋白以及来自健康供体心脏（对照）的心肌蛋白。分析了10个来自扩张型心肌病（DCM）心脏移植的RA凝胶与10个LV凝胶，以寻找其斑点模式的定量差异。在与每个凝胶匹配的197个斑点中，DCM患者的RA和LV之间有40个斑点的强度存在显著差异。分析了更多来自DCM患者和健康供体心脏的心房和心室凝胶（20个RA凝胶，20个LV凝胶；4个RA凝胶，4个LV凝胶），以寻找蛋白质表达的定性差异。DCM患者所有RA凝胶中存在的三个蛋白质斑点（SSP 1120：分子量/等电点：20.5 kDa/4.6；SSP 1119：分子量/等电点：20.6 kDa/4.5；SSP 0117：分子量/等电点：20.7/<4.5）在所有LV凝胶中均不存在。两个蛋白质斑点（SSP 0112：分子量/等电点：17.2 kDa，/<4.4；SSP 0114：分子量/等电点：17.6 kDa/<4.4）仅出现在所有LV凝胶中，而不出现在RA凝胶中。这五个定性不同的斑点在DCM患者和健康供体心脏中是相同的。对一些不同的斑点进行了内部测序，并分别鉴定为肌球蛋白轻链同工型（心房肌球蛋白轻链2；心室肌球蛋白轻链2；心房肌球蛋白轻链1），其蛋白质识别资源（PIR）登录号分别为A44451、S03708、A30881。此外，还鉴定出磷酸甘油酸变位酶（PIR：JQ0750）和ATP合酶α链（PIR：S17193）。因此，通过二维聚丙烯酰胺凝胶电泳确定了心房和心室蛋白质模式在定量和定性方面的差异。使用我们的方法发现了人心房和心室心肌中肌球蛋白轻链的特征性分布。
