Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, P. R. China.
Proteomics. 2011 Apr;11(8):1473-85. doi: 10.1002/pmic.201000482. Epub 2011 Feb 25.
Salvianolic acid B (SB) is a natural compound with protective effect against ischemia-reperfusion heart injury. However, the signal network of SB including both direct target proteins and downstream signal-related proteins has not been clarified. In the present study, epidermal growth factor receptor (EGFR) was predicted to be the most possible direct protein target of SB by INVDOCK, a ligand-protein inverse-docking algorithm. Possible signal-related proteins of SB in H9C2 cells, including both under normal condition and under ischemia-reperfusion injury, were searched using 2-DE analysis. Totally, 14 signal-related proteins were found. Finally, signal network from EGFR to the signal-related proteins was established using bioinformatic analysis. Interestingly, 9 of the 14 signal-related proteins could be included in a network together with EGFR through direct interaction or only one intermediate partner. The signal cascade from EGFR to heat shock protein 27 (HSP27) and mitofilin (IMMT, inner membrane mitochondrial protein) might be the most important cascade. The signal network was certified by measuring the binding affinity of SB to EGFR in vitro, the effect of SB on internalization and phosphorylation of EGFR, the effect of SB on viability and proliferation of H9C2 cells, and the expression of inner membrane mitochondrial protein in the presence of EGFR inhibitor AG 1478.
丹酚酸 B（SB）是一种具有抗缺血再灌注心脏损伤作用的天然化合物。然而，SB 的信号网络，包括直接靶蛋白和下游信号相关蛋白，尚未阐明。在本研究中，通过配体-蛋白反向对接算法 INVDOCK 预测表皮生长因子受体（EGFR）是 SB 最可能的直接靶蛋白。使用 2-DE 分析，在正常和缺血再灌注损伤条件下，在 H9C2 细胞中搜索 SB 的可能信号相关蛋白。总共发现了 14 种信号相关蛋白。最后，使用生物信息学分析建立了从 EGFR 到信号相关蛋白的信号网络。有趣的是，通过直接相互作用或仅一个中间伴侣，这 14 种信号相关蛋白中的 9 种可以与 EGFR 一起包含在一个网络中。从 EGFR 到热休克蛋白 27（HSP27）和线粒体内膜蛋白（IMMT）的信号级联可能是最重要的级联。通过体外测量 SB 与 EGFR 的结合亲和力、SB 对内化和 EGFR 磷酸化的影响、SB 对 H9C2 细胞活力和增殖的影响以及在 EGFR 抑制剂 AG 1478 存在下内膜线粒体蛋白的表达，验证了信号网络。
