Flinkman Dani, Hong Ye, Gnjatovic Jelena, Deshpande Prasannakumar, Ortutay Zsuzsanna, Peltonen Sirkku, Kaasinen Valtteri, James Peter, Coffey Eleanor
Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.
Lund University, Department of Immunotechnology, Lund, Sweden.
NPJ Parkinsons Dis. 2023 Feb 6;9(1):20. doi: 10.1038/s41531-023-00460-w.
Deficits in protein synthesis are associated with Parkinson's disease (PD). However, it is not known which proteins are affected or if there are synthesis differences between patients with sporadic and Leucine-Rich Repeat Kinase 2 (LRRK2) G2019S PD, the most common monogenic form. Here we used bio-orthogonal non-canonical amino acid tagging for global analysis of newly translated proteins in fibroblasts from sporadic and LRKK2-G2019S patients. Quantitative proteomic analysis revealed that several nascent proteins were reduced in PD samples compared to healthy without any significant change in mRNA levels. Using targeted proteomics, we validated which of these proteins remained dysregulated at the static proteome level and found that regulators of endo-lysosomal sorting, mRNA processing and components of the translation machinery remained low. These proteins included autophagy-related protein 9A (ATG9A) and translational stability regulator YTH N6-ethyladenosine RNA binding protein 3 (YTHDF3). Notably, 77% of the affected proteins in sporadic patients were also repressed in LRRK2-G2019S patients (False discovery rate (FDR) < 0.05) in both sporadic and LRRK2-G2019S samples. This analysis of nascent proteomes from PD patient skin cells reveals that regulators of proteostasis are repressed in both sporadic and LRRK2-G2019S PD.
蛋白质合成缺陷与帕金森病（PD）相关。然而，尚不清楚哪些蛋白质受到影响，也不清楚散发性患者与最常见的单基因形式——富含亮氨酸重复激酶2（LRRK2）G2019S型帕金森病患者之间是否存在合成差异。在这里，我们使用生物正交非标准氨基酸标记技术对散发性和LRRK2 - G2019S型患者成纤维细胞中新翻译的蛋白质进行全局分析。定量蛋白质组学分析显示，与健康对照相比，PD样本中的几种新生蛋白质减少，而mRNA水平没有任何显著变化。使用靶向蛋白质组学，我们验证了哪些蛋白质在静态蛋白质组水平上仍然失调，并发现内溶酶体分选调节因子、mRNA加工以及翻译机制的组成部分仍然减少。这些蛋白质包括自噬相关蛋白9A（ATG9A）和翻译稳定性调节因子YTH N6 - 乙基腺苷RNA结合蛋白3（YTHDF3）。值得注意的是，在散发性和LRRK2 - G2019S样本中，散发性患者中77%受影响蛋白质在LRRK2 - G2019S患者中也受到抑制（错误发现率（FDR）<0.05）。对PD患者皮肤细胞新生蛋白质组的分析表明，蛋白质稳态调节因子在散发性和LRRK2 - G2019S型帕金森病中均受到抑制。
