MRS2 missense variation at Asp216 abrogates inhibitory Mg2+ binding, potentiating cell migration and apoptosis resistance.

IF 4.5 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Protein Science Pub Date : 2024-08-01 DOI:10.1002/pro.5108
Sukanthathulse Uthayabalan, Taylor Lake, Peter B Stathopulos
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Abstract

Mitochondrial magnesium (Mg2+) is a crucial modulator of protein stability, enzymatic activity, ATP synthesis, and cell death. Mitochondrial RNA splicing protein 2 (MRS2) is the main Mg2+ channel in the inner mitochondrial membrane that mediates influx into the matrix. Recent cryo-electron microscopy (cryo-EM) human MRS2 structures exhibit minimal conformational changes at high and low Mg2+, yet the regulation of human MRS2 and orthologues by Mg2+ binding to analogous matrix domains has been well established. Further, a missense variation at D216 has been identified associated with malignant melanoma and MRS2 expression and activity is implicated in gastric cancer. Thus, to gain more mechanistic and functional insight into Mg2+ sensing by the human MRS2 matrix domain and the association with proliferative disease, we assessed the structural, biophysical, and functional effects of a D216Q mutant. We show that the D216Q mutation is sufficient to abrogate Mg2+-binding and associated conformational changes including increased α-helicity, stability, and monomerization. Further, we reveal that the MRS2 matrix domains interact with ~μM affinity, which is weakened by up to two orders of magnitude in the presence of Mg2+ for wild-type but unaffected for D216Q. Finally, we demonstrate the importance of Mg2+ sensing by MRS2 to prevent matrix Mg2+ overload as HeLa cells overexpressing MRS2 show enhanced Mg2+ uptake, cell migration, and resistance to apoptosis while MRS2 D216Q robustly potentiates these cancer phenotypes. Collectively, our findings further define the MRS2 matrix domain as a critical Mg2+ sensor that undergoes conformational and assembly changes upon Mg2+ interactions dependent on D216 to temper matrix Mg2+ overload.

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MRS2在Asp216处的错义变异削弱了Mg2+的抑制性结合,从而增强了细胞迁移和抗凋亡能力。
线粒体镁(Mg2+)是蛋白质稳定性、酶活性、ATP 合成和细胞死亡的重要调节因子。线粒体 RNA 剪接蛋白 2(MRS2)是线粒体内膜上主要的 Mg2+ 通道,它介导 Mg2+ 流入基质。最新的低温电子显微镜(cryo-EM)人类 MRS2 结构在高 Mg2+ 和低 Mg2+ 时的构象变化极小,但人类 MRS2 和同源物通过 Mg2+ 与类似基质结构域的结合进行调控的特性已得到证实。此外,还发现 D216 的错义变异与恶性黑色素瘤有关,MRS2 的表达和活性与胃癌有关。因此,为了从机理和功能上更深入地了解人类 MRS2 基质结构域对 Mg2+ 的感应以及与增殖性疾病的关联,我们评估了 D216Q 突变体的结构、生物物理和功能效应。我们发现 D216Q 突变足以导致 Mg2+ 结合失效以及相关的构象变化,包括α-异构性、稳定性和单聚化的增加。此外,我们还发现 MRS2 矩阵结构域以 ~μM 的亲和力相互作用,在 Mg2+ 存在的情况下,野生型的亲和力最多会减弱两个数量级,而 D216Q 则不受影响。最后,我们证明了MRS2感知Mg2+对防止基质Mg2+超载的重要性,因为过表达MRS2的HeLa细胞表现出更强的Mg2+吸收、细胞迁移和抗凋亡能力,而MRS2 D216Q则显著增强了这些癌症表型。总之,我们的研究结果进一步确定了MRS2基质结构域是一个关键的Mg2+传感器,它在依赖于D216的Mg2+相互作用时发生构象和组装变化,以调节基质Mg2+过载。
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来源期刊
Protein Science
Protein Science 生物-生化与分子生物学
CiteScore
12.40
自引率
1.20%
发文量
246
审稿时长
1 months
期刊介绍: Protein Science, the flagship journal of The Protein Society, is a publication that focuses on advancing fundamental knowledge in the field of protein molecules. The journal welcomes original reports and review articles that contribute to our understanding of protein function, structure, folding, design, and evolution. Additionally, Protein Science encourages papers that explore the applications of protein science in various areas such as therapeutics, protein-based biomaterials, bionanotechnology, synthetic biology, and bioelectronics. The journal accepts manuscript submissions in any suitable format for review, with the requirement of converting the manuscript to journal-style format only upon acceptance for publication. Protein Science is indexed and abstracted in numerous databases, including the Agricultural & Environmental Science Database (ProQuest), Biological Science Database (ProQuest), CAS: Chemical Abstracts Service (ACS), Embase (Elsevier), Health & Medical Collection (ProQuest), Health Research Premium Collection (ProQuest), Materials Science & Engineering Database (ProQuest), MEDLINE/PubMed (NLM), Natural Science Collection (ProQuest), and SciTech Premium Collection (ProQuest).
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