Barallobre-Barreiro Javier, Gupta Shashi K, Zoccarato Anna, Kitazume-Taneike Rika, Fava Marika, Yin Xiaoke, Werner Tessa, Hirt Marc N, Zampetaki Anna, Viviano Alessandro, Chong Mei, Bern Marshall, Kourliouros Antonios, Domenech Nieves, Willeit Peter, Shah Ajay M, Jahangiri Marjan, Schaefer Liliana, Fischer Jens W, Iozzo Renato V, Viner Rosa, Thum Thomas, Heineke Joerg, Kichler Antoine, Otsu Kinya, Mayr Manuel
From King's British Heart Foundation Centre, King's College London, United Kingdom (J.B.-B., A. Zoccarato, R.K.-T., M.F., X.Y., A. Zampetaki, M.C., P.W., A.M.S., K.O., M.M.); Institute for Molecular and Translational Therapeutic Strategies, MH-Hannover, Germany (S.K.G., T.T.); St George's Hospital, NHS Trust, London, United Kingdom (M.F., A.V., A.K., M.J.); University Medical Center Hamburg-Eppendorf, Germany (T.W., M.N.H.); Protein Metrics, San Carlos, CA (M.B.); Biobanco A Coruña, INIBIC-Complexo Hospitalario Universitario de A Coruña, Spain (N.D.); Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany (L.S.); Institute for Pharmacology and Clinical Pharmacology, Heinrich-Heine-University, Düsseldorf, Germany (J.W.F.); Sidney Kimmel Medical College at Thomas Jefferson University, Philadelphia, PA (R.V.I.); Thermo Fisher Scientific, San Jose, CA (R.V.); Experimental Cardiology, Department of Cardiology and Angiology, MH-Hannover, Germany (J.H.); and Laboratoire Vecteurs: Synthèse et Applications Thérapeutiques, UMR 7199 CNRS Université de Strasbourg, Illkirch, France (A.K.).
Circulation. 2016 Sep 13;134(11):817-32. doi: 10.1161/CIRCULATIONAHA.115.016423. Epub 2016 Aug 24.
Myocardial fibrosis is a feature of many cardiac diseases. We used proteomics to profile glycoproteins in the human cardiac extracellular matrix (ECM).
Atrial specimens were analyzed by mass spectrometry after extraction of ECM proteins and enrichment for glycoproteins or glycopeptides.
ECM-related glycoproteins were identified in left and right atrial appendages from the same patients. Several known glycosylation sites were confirmed. In addition, putative and novel glycosylation sites were detected. On enrichment for glycoproteins, peptides of the small leucine-rich proteoglycan decorin were identified consistently in the flowthrough. Of all ECM proteins identified, decorin was found to be the most fragmented. Within its protein core, 18 different cleavage sites were identified. In contrast, less cleavage was observed for biglycan, the most closely related proteoglycan. Decorin processing differed between human ventricles and atria and was altered in disease. The C-terminus of decorin, important for the interaction with connective tissue growth factor, was detected predominantly in ventricles in comparison with atria. In contrast, atrial appendages from patients in persistent atrial fibrillation had greater levels of full-length decorin but also harbored a cleavage site that was not found in atrial appendages from patients in sinus rhythm. This cleavage site preceded the N-terminal domain of decorin that controls muscle growth by altering the binding capacity for myostatin. Myostatin expression was decreased in atrial appendages of patients with persistent atrial fibrillation and hearts of decorin null mice. A synthetic peptide corresponding to this decorin region dose-dependently inhibited the response to myostatin in cardiomyocytes and in perfused mouse hearts.
This proteomics study is the first to analyze the human cardiac ECM. Novel processed forms of decorin protein core, uncovered in human atrial appendages, can regulate the local bioavailability of antihypertrophic and profibrotic growth factors.
心肌纤维化是许多心脏疾病的一个特征。我们运用蛋白质组学技术对人心脏细胞外基质(ECM)中的糖蛋白进行了分析。
在提取ECM蛋白并富集糖蛋白或糖肽后,通过质谱分析法对心房标本进行分析。
在同一患者的左、右心耳中鉴定出了与ECM相关的糖蛋白。证实了几个已知的糖基化位点。此外,还检测到了推定的和新的糖基化位点。在富集糖蛋白时,始终在流出物中鉴定出富含亮氨酸的小分子蛋白聚糖核心蛋白聚糖的肽段。在鉴定出的所有ECM蛋白中,核心蛋白聚糖是片段化程度最高的。在其蛋白核心内,鉴定出了18个不同的切割位点。相比之下,与核心蛋白聚糖关系最密切的蛋白聚糖双糖链蛋白聚糖的切割较少。核心蛋白聚糖的加工在人的心室和心房之间存在差异,并且在疾病中会发生改变。与心房相比,核心蛋白聚糖对结缔组织生长因子相互作用很重要的C末端主要在心室中检测到。相比之下,持续性房颤患者的心耳中全长核心蛋白聚糖水平更高,但也有一个在窦性心律患者的心耳中未发现的切割位点。这个切割位点位于核心蛋白聚糖的N末端结构域之前,该结构域通过改变对肌肉生长抑制素的结合能力来控制肌肉生长。持续性房颤患者的心耳和核心蛋白聚糖基因敲除小鼠的心脏中肌肉生长抑制素表达降低。与该核心蛋白聚糖区域对应的合成肽可剂量依赖性地抑制心肌细胞和灌注小鼠心脏中对肌肉生长抑制素的反应。
这项蛋白质组学研究首次对人心脏ECM进行了分析。在人心脏心耳中发现的核心蛋白聚糖蛋白核心的新加工形式可调节抗肥厚和促纤维化生长因子的局部生物利用度。