Phosphoproteomic analysis identifies phospho-Threonine-17 site of phospholamban important in low molecular weight isoform of fibroblast growth factor 2-induced protection against post-ischemic cardiac dysfunction.
Phosphoproteomic analysis identifies phospho-Threonine-17 site of phospholamban important in low molecular weight isoform of fibroblast growth factor 2-induced protection against post-ischemic cardiac dysfunction.
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磷蛋白组学分析确定了磷蛋白的磷苏氨酸-17位点在成纤维细胞生长因子2诱导的保护缺血性心功能障碍的低分子量异构体中起重要作用。
DOI：
10.1016/j.yjmcc.2020.08.006
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发表时间：
2020-11
影响因子：
5
通讯作者：
Schultz JEJ
中科院分区：
文献类型：
--
作者：
Manning JR;Wijeratne AB;Oloizia BB;Zhang Y;Greis KD;Schultz JEJ
成纤维细胞生长因子 2 (FGF2) 具有多种生物学作用，其中之一是保护心脏免受缺血性发作相关的功能障碍和损伤。我们的实验室证明，FGF2 的低分子量 (LMW) 亚型可防止心肌功能障碍，而高分子量 (HMW) 亚型则与缺血后心脏功能恢复的恶化有关。 LMW FGF2 介导的心脏保护作用是通过激活多种激酶（包括 PKCalpha、PKCepsilon 和 ERK）以及抑制 p38 和 JNK 来促进的。然而，与 LMW FGF2 诱导的针对心肌功能障碍的心脏保护作用相关的那些激酶的底物仍有待阐明。为了确定 LMW FGF2 改善缺血后心脏功能的底物，对仅表达 LMW FGF2 的小鼠心脏进行缺血再灌注 (I/R) 损伤，并通过基于质谱 (MS) 的定量磷酸化蛋白质组策略进行分析。 MS 分析确定了 7 种肌内质网 (SR) 蛋白的 50 个磷酸化位点，与缺乏 FGF2 的心脏相比，仅表达 LMW FGF2 的 I/R 处理心脏中这些位点发生了显着改变。已鉴定的磷酸化 SR 蛋白之一是受磷蛋白 (PLB)，在仅表达 LMW FGF2 的心脏 I/R 后，其在苏氨酸 17 (Thr17) 处表现出快速、增加的磷酸化；使用基于单反应监测的 MS 工作流程进一步验证了这一点。为了证明磷酸化 Thr17 PLB 在 LMW FGF2 介导的心脏保护中的机制作用，对仅表达 LMW FGF2 的心脏和仅表达 LMW FGF2 且具有缺乏可磷酸化 Thr17 的突变 PLB (Thr17Ala PLB) 的心脏进行 I/R。仅表达LMW FGF2的心脏显示I/R后心脏功能的恢复显着改善(p<0.05)，并且这种功能改善在表达LMW FGF2和Thr17Ala PLB的心脏中显着消除(p<0.05)。研究结果表明，LMW FGF2 通过对 I/R 损伤恢复至关重要的 SR 蛋白的调节变化来调节细胞内钙处理/循环，从而保护心脏免受缺血后心功能障碍的影响。
Among its many biological roles, fibroblast growth factor 2 (FGF2) protects the heart from dysfunction and damage associated with an ischemic attack. Our laboratory demonstrated that its protection against myocardial dysfunction occurs by the low molecular weight (LMW) isoform of FGF2, while the high molecular weight (HMW) isoforms are associated with a worsening in post-ischemic recovery of cardiac function. LMW FGF2-mediated cardioprotection is facilitated by activation of multiple kinases, including PKCalpha, PKCepsilon, and ERK, and inhibition of p38 and JNK. Yet, the substrates of those kinases associated with LMW FGF2-induced cardioprotection against myocardial dysfunction remain to be elucidated. To identify substrates in LMW FGF2 improvement of post-ischemic cardiac function, mouse hearts expressing only LMW FGF2 were subjected to ischemia-reperfusion (I/R) injury and analyzed by a mass spectrometry (MS)-based quantitative phosphoproteomic strategy. MS analysis identified 50 phosphorylation sites from 7 sarcoendoplasmic reticulum (SR) proteins that were significantly altered in I/R-treated hearts only expressing LMW FGF2 compared to those hearts lacking FGF2. One of those phosphorylated SR proteins identified was phospholamban (PLB), which exhibited rapid, increased phosphorylation at Threonine-17 (Thr17) after I/R in hearts expressing only LMW FGF2; this was further validated using Single Reaction Monitoring-based MS workflow. To demonstrate a mechanistic role of phospho-Thr17 PLB in LMW FGF2-mediated cardioprotection, hearts only expressing LMW FGF2 and those expressing only LMW FGF2 with a mutant PLB lacking phosphorylatable Thr17 (Thr17Ala PLB) were subjected to I/R. Hearts only expressing LMW FGF2 showed significantly improved recovery of cardiac function following I/R (p<0.05), and this functional improvement was significantly abrogated in hearts expressing LMW FGF2 and Thr17Ala PLB (p<0.05). The findings indicate that LMW FGF2 modulates intracellular calcium handling/cycling via regulatory changes in SR proteins essential for recovery from I/R injury, and thereby protects the heart from post-ischemic cardiac dysfunction.
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影响因子：
5
作者：
Gabel, Scott A;Walker, Vickie R;Murphy, Elizabeth
通讯作者：
Murphy, Elizabeth
DOI：
10.3109/08977194.2012.656759
发表时间：
2012-04
期刊：
Growth factors (Chur, Switzerland)
影响因子：
--
作者：
Manning JR;Carpenter G;Porter DR;House SL;Pietras DA;Doetschman T;Schultz Jel J
通讯作者：
Schultz Jel J
影响因子：
5.6
作者：
Bousette, Nicolas;Abbasi, Cynthia;Gramolini, Anthony O.
通讯作者：
Gramolini, Anthony O.
影响因子：
5.6
作者：
Dini, G;Funghini, S;Del Rosso, M
通讯作者：
Del Rosso, M
FGF12 是 Brugada 综合征的候选基因座。
影响因子：
5.5
作者：
Hennessey, Jessica A.;Marcou, Cherisse A.;Wang, Chuan;Wei, Eric Q.;Wang, Chaojian;Tester, David J.;Torchio, Margherita;Dagradi, Federica;Crotti, Lia;Schwartz, Peter J.;Ackerman, Michael J.;Pitt, Geoffrey S.
通讯作者：
Pitt, Geoffrey S.