Aims Prolyl hydroxylation is a post-translational modification that regulates protein stability, turnover, and activity. The proteins that catalyze prolyl hydroxylation belong to the 2-oxoglutarate- and iron-dependent oxygenase family of proteins. 2-oxoglutarate- and iron-dependent oxygenase domain-containing protein 1 (Ogfod1), which hydroxylates a proline in ribosomal protein s23 is a newly described member of this family. The aims of this study were to investigate roles for Ogfod1 in the heart, and in the heart’s response to stress. Methods and results We isolated hearts from wild-type (WT) and Ogfod1 knockout (KO) mice and performed quantitative proteomics using tandem mass Tag labelling coupled to liquid chromatography and tandem mass spectrometry (LC-MS/MS) to identify protein changes. Ingenuity pathway analysis identified ‘Urate Biosynthesis/Inosine 5′-phosphate Degradation’ and ‘Purine Nucleotides Degradation II (Aerobic)’ as the most significantly enriched pathways. We performed metabolomics analysis and found that both purine and pyrimidine pathways were altered with the purine nucleotide inosine 5′-monophosphate showing a 3.5-fold enrichment in KO hearts (P = 0.011) and the pyrimidine catabolism product beta-alanine showing a 1.7-fold enrichment in KO hearts (P = 0.014). As changes in these pathways have been shown to contribute to cardioprotection, we subjected isolated perfused hearts to ischaemia and reperfusion (I/R). KO hearts showed a 41.4% decrease in infarct size and a 34% improvement in cardiac function compared to WT hearts. This protection was also evident in an in vivo I/R model. Additionally, our data show that treating isolated perfused WT hearts with carnosine, a metabolite of beta-alanine, improved protection in the context of I/R injury, whereas treating KO hearts with carnosine had no impact on recovery of function or infarct size. Conclusions Taken together, these data show that Ogfod1 deletion alters the myocardial proteome and metabolome to confer protection against I/R injury.

中文翻译：

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Ogfod1 缺失增加心脏β-丙氨酸水平并保护小鼠免受缺血再灌注损伤

目的脯氨酰羟基化是一种翻译后修饰，可调节蛋白质的稳定性、周转率和活性。催化脯氨酰羟基化的蛋白质属于蛋白质的 2-氧戊二酸和铁依赖性加氧酶家族。2-氧代戊二酸和铁依赖的含氧酶结构域蛋白 1 (Ogfod1)，它使核糖体蛋白 s23 中的脯氨酸羟基化，是该家族的新成员。本研究的目的是调查 Ogfod1 在心脏中的作用，以及心脏对压力的反应。方法和结果 我们从野生型 (WT) 和 Ogfod1 敲除 (KO) 小鼠中分离出心脏，并使用串联质量标签标记结合液相色谱和串联质谱 (LC-MS/MS) 进行定量蛋白质组学以识别蛋白质变化。Ingenuity 途径分析确定“尿酸盐生物合成/肌苷 5'-磷酸降解”和“嘌呤核苷酸降解 II（需氧）”是最显着富集的途径。我们进行了代谢组学分析，发现嘌呤和嘧啶途径都发生了改变，嘌呤核苷酸肌苷 5'-单磷酸在 KO 心脏中显示出 3.5 倍的富集（P = 0.011），而嘧啶分解代谢产物β-丙氨酸显示出 1.7 倍KO 心脏中的富集 (P = 0.014)。由于这些途径的变化已被证明有助于心脏保护，我们对孤立的灌注心脏进行缺血和再灌注 (I/R)。与 WT 心脏相比，KO 心脏的梗死面积减少了 41.4%，心脏功能改善了 34%。这种保护在体内 I/R 模型中也很明显。此外，我们的数据表明，用肌肽（一种 β-丙氨酸的代谢物）治疗孤立的灌注 WT 心脏，在 I/R 损伤的情况下改善了保护，而用肌肽治疗 KO 心脏对功能恢复或梗塞面积没有影响。结论 综上所述，这些数据表明 Ogfod1 缺失改变了心肌蛋白质组和代谢组，从而提供了对 I/R 损伤的保护。