Cardiovascular Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Animal Surgery and Resources Core Facility, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Cardiovasc Res. 2022 Oct 21;118(13):2847-2858. doi: 10.1093/cvr/cvab323.
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.
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.
Taken together, these data show that Ogfod1 deletion alters the myocardial proteome and metabolome to confer protection against I/R injury.
脯氨酰羟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。催化脯氨酰羟化的蛋白质属于 2-酮戊二酸和铁依赖性氧合酶家族的蛋白质。2-酮戊二酸和铁依赖性氧合酶结构域包含蛋白 1（Ogfod1），它羟化核糖体蛋白 s23 中的一个脯氨酸，是该家族的一个新描述的成员。本研究的目的是研究 Ogfod1 在心脏中的作用，以及在心脏对压力的反应中的作用。
我们从野生型（WT）和 Ogfod1 敲除（KO）小鼠中分离心脏，并使用串联质量标签标记与液相色谱和串联质谱（LC-MS/MS）耦合的定量蛋白质组学来鉴定蛋白质变化。Ingenuity 通路分析将“尿酸合成/肌苷 5'-磷酸降解”和“嘌呤核苷酸降解 II（需氧）”鉴定为最显著富集的通路。我们进行代谢组学分析，发现嘌呤和嘧啶途径都发生了改变，在 KO 心脏中，嘌呤核苷酸肌苷 5'-单磷酸（P = 0.011）富集了 3.5 倍，嘧啶分解产物 β-丙氨酸（P = 0.014）富集了 1.7 倍。由于这些途径的变化已被证明有助于心脏保护，我们将分离的灌注心脏进行缺血再灌注（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%。在体内 I/R 模型中也观察到了这种保护作用。此外，我们的数据表明，用β-丙氨酸的代谢产物肌肽处理分离的灌注 WT 心脏可改善 I/R 损伤情况下的保护作用，而用肌肽处理 KO 心脏对功能恢复或梗死面积没有影响。
总之，这些数据表明，Ogfod1 缺失改变了心肌蛋白质组和代谢组，从而对 I/R 损伤提供保护。
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