Hou Yanwen, Le Bihan Marie-Catherine, Vega-Avelaira David, Coulton Gary R
Division Cardiac & Vascular Sciences and Division Basic Medical Sciences, St. George's, University of London, Cranmer Terrace, London, UK.
Proteomics. 2006 May;6(10):3096-108. doi: 10.1002/pmic.200500475.
Complex molecular changes associated with early stage human heart disease are poorly understood and prevent the development of effective treatments of human cardiac disease. Relatively minor structural changes in early disease may accompany some conditions such as arrhythmias. Our objective was to determine if significant proteomic changes occur in heart tissues in the absence of structural pathology. We used a proteomic "pipeline" based on Ciphergen SELDI-TOF/MS, gel electrophoresis and MALDI-TOF/MS. The kyphoscoliosis (ky) mouse carries a mutation in a putative transglutaminase causing a primary skeletal muscle disease. The ky protein is expressed usually in skeletal and cardiac muscle but its absence from the ky heart causes no structural pathology making it a good model of "occult" heart disease. We discovered 20 statistically validated biomarkers discriminating ky from normal hearts, one cardiac troponin-I was reduced by 40% in ky hearts. A 17% deficit was confirmed subsequently by Western blot. Thus, the proteome of ky hearts was abnormal, giving support to our contention that this SELDI-based analytical approach is capable of making a significant contribution to the analysis of complex proteomic changes in early stage human heart disease.
与早期人类心脏病相关的复杂分子变化目前仍知之甚少，这阻碍了人类心脏病有效治疗方法的开发。早期疾病中相对较小的结构变化可能伴随着某些病症，如心律失常。我们的目标是确定在没有结构病理学的情况下，心脏组织中是否会发生显著的蛋白质组学变化。我们使用了一种基于Ciphergen表面增强激光解吸电离飞行时间质谱（SELDI-TOF/MS）、凝胶电泳和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/MS）的蛋白质组学“流程”。脊柱后凸（ky）小鼠在一种假定的转谷氨酰胺酶中发生突变，导致原发性骨骼肌疾病。ky蛋白通常在骨骼肌和心肌中表达，但ky心脏中缺乏该蛋白不会导致结构病理学变化，使其成为“隐匿性”心脏病的良好模型。我们发现了20种经统计学验证的可区分ky心脏与正常心脏的生物标志物，其中一种心肌肌钙蛋白I在ky心脏中减少了40%。随后通过蛋白质印迹法证实了17%的缺陷。因此，ky心脏的蛋白质组是异常的，这支持了我们的观点，即这种基于SELDI的分析方法能够对早期人类心脏病复杂蛋白质组学变化的分析做出重大贡献。
