Activated mitochondrial dynamin-related protein 1 couples with LRRK2 to cause mitochondrial dysfunction in hypoxic cells

Chenyang DUAN; Xinming XIANG; Lei KUANG; Liangming LIU; Tao LI

doi:10.16016/j.1000-5404.201910153

您当前的位置：

首页 >

文章列表页 >

Activated mitochondrial dynamin-related protein 1 couples with LRRK2 to cause mitochondrial dysfunction in hypoxic cells

Basic Medicine | 更新时间：2026-01-09

- Activated mitochondrial dynamin-related protein 1 couples with LRRK2 to cause mitochondrial dysfunction in hypoxic cells
- Journal of Third Military Medical University Vol. 42, Issue 4, Pages: 326-333(2020)
- 作者机构：
  
  400042 重庆，陆军军医大学(第三军医大学)野战外科研究所战伤休克与输血研究室，创伤、烧伤与复合伤国家重点实验室
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81730059);the National Natural Science Foundation for Young Scholars of China(81700429)
- DOI：10.16016/j.1000-5404.201910153
  CLC： R341; R363.21; R364.4
- Received：22 October 2019，
  
  Revised：2019-12-02，
  
  Published：28 February 2020
- 稿件说明：
移动端阅览
Chenyang DUAN, Xinming XIANG, Lei KUANG, et al. Activated mitochondrial dynamin-related protein 1 couples with LRRK2 to cause mitochondrial dysfunction in hypoxic cells[J]. Journal of Third Military Medical University, 2020, 42(4): 326-333.
DOI：

Chenyang DUAN, Xinming XIANG, Lei KUANG, et al. Activated mitochondrial dynamin-related protein 1 couples with LRRK2 to cause mitochondrial dysfunction in hypoxic cells[J]. Journal of Third Military Medical University, 2020, 42(4): 326-333. DOI： 10.16016/j.1000-5404.201910153.

摘要

目的

探究缺氧后活化Drp1对线粒体能量代谢和有氧呼吸的调控机制。

方法

在心肌细胞(H9C2)、血管平滑肌细胞系(vascular smooth muscle cells，VSMC)、肠上皮细胞系(intestinal epithelial cells，IEC)中通过免疫共沉淀、免疫荧光方法观察多种细胞类型缺氧后Drp1活性变化及与线粒体损伤的关系；利用ZDOCK方法建立Drp1与同源激酶LRRK2蛋白互作模型，验证缺氧后Drp1-LRRK2偶联情况；以VSMC细胞为代表，通过Drp1点突变T595A破坏缺氧后Drp1-LRRK2的偶联，分别检测了培养基酸化程度、细胞外乳酸含量等反映无氧呼吸的指标和线粒体膜电位、ATP生成量等反映有氧呼吸的指标。

结果

缺氧后Drp1的Thr磷酸化水平增高、线粒体损伤明显。进一步研究发现缺氧后Drp1蛋白与LRRK2蛋白界面残基间存在大量氢键，导致缺氧后Drp1-LRRK2偶联从而影响线粒体功能。具体表现为糖酵解过程中乳酸生成的增多和线粒体有氧呼吸过程中膜电位的减低。

结论

缺氧后活化Drp1可通过偶联LRRK2破坏线粒体能量代谢和有氧呼吸，加重多器官组织线粒体损伤。

Abstract

Objective

To explore the mechanism of activated mitochondrial dynamin-related protein 1 (Drp1) in regulating mitochondrial energy metabolism and aerobic respiration after hypoxia.

Methods

We observed the changes of Drp1 activity and its relationship with mitochondrial damage by coimmunoprecipitation and immunofluorescence in cardiac myocyte line H9C2

vascular smooth muscle cells (VSMCs) and intestinal epithelial cells (IECs). The protein interaction model of Drp1 and LRRK2 was established using ZDOCK and verified by coimmunoprecipitation. A point mutation (T595A) was introduced in Drp1 gene to disrupt the coupling of Drp1 and LRRK2 in VSMCs

and the acidification of the culture medium

extracellular lactate content

mitochondrial membrane potential and ATP production of the cells were examined.

Results

Hypoxia caused significantly increased level of Thr phosphorylation in Drp1 and obvious mitochondrial damage in the cells. After hypoxia

numerus hydrogen bonds were found between the interface residues of Drp1 and LRRK2 proteins

which resulted in close binding between them to cause mitochondrial dysfunction

manifested by an increased lactate production during glycolysis and a decreased mitochondrial membrane potential during mitochondrial aerobic respiration.

Conclusion

Activated Drp1 causes impairment of mitochondrial energy metabolism and aerobic respiration by coupling with LRRK2 to aggravate mitochondrial damage in multiple tissues and organs after hypoxia.

关键词

Keywords

references

DUAN C Y, CAO Z Z, TANG F Q, et al . MiRNA-mRNA crosstalk in myocardial ischemia induced by calcified aortic valve stenosis[J ] . Aging (Albany NY), 2019, 11(2): 448-466. DOI: 10.18632/aging.101751.

LIU Z Q, ZENG X, DUAN C Y. Neuropsychological rehabilitation and psychotherapy of adult traumatic brain injury patients with depression: a systematic review and meta-analysis[J]. J Neurosurg Sci, 2018, 62(1): 24-35. DOI: 10.23736/S0390-5616.17.03953-4.

DUAN C Y, CHEN K, YANG G M, et al . HIF-1α regulates Cx40-dependent vasodilatation following hemorrhagic shock in rats[J ] . Am J Transl Res, 2017, 9(3): 1277-1286.

DUAN C Y, YANG G M, LI T, et al . Advances in vascularhyporeactivity after shoc k: the mechanisms and managements[J ] . Shock, 2015, 44(6): 524-534. DOI: 10.1097/SHK.0000000000000457.

MEI Z B, DUAN C Y, LI C B, et al . Prognostic role of tumor PIK3CA mutation in colorectal cancer: a systematic review and meta-analysis[J ] . Ann Oncol, 2016, 27(10): 1836-1848. DOI: 10.1093/annonc/mdw264.

FUHRMANN D C, BRÜNE B. Mitochondrial composition and function under the control of hypoxia[J]. Redox Biol, 2017, 12: 208-215. DOI: 10.1016/j.redox.2017.02.012.

MARSBOOM G, TOTH P T, RYAN J J, et al . Dynamin-related protein 1-mediated mitochondrial mitotic fission permits hyperproliferation of vascular smooth muscle cells and offers a novel therapeutic target in pulmonary hypertension[J ] . Circ Res, 2012, 110(11): 1484-1497. DOI: 10.1161/CIRCRESAHA.111.263848.

NISHIMURA A, SHIMAUCHI T, TANAKA T, et al . Hypoxia- induced interaction offilamin with Drp1 causes mitochondrial hyperfission-associated myocardial senescence[J ] . Sci Signal, 2018, 11(556): eaat5185. DOI: 10.1126/scisignal.aat5185.

WANG X L, YAN M H, FUJIOKA H, et al . LRRK2 regulates mitochondrial dynamic s and function through direct interaction with DLP1[J ] . Hum Mol Genet, 2012, 21(9): 1931-1944. DOI: 10.1093/hmg/dds003.

DILIP A, LEŠNIK S, ŠTULAR T, et al . Ligand-based virtual screening interface between PyMOL and LiSiCA[J ] . J Cheminform, 2016, 8(1): 46. DOI: 10.1186/s13321-016-0157-z.

PIERCE B G, HOURAI Y, WENG Z P. Accelerating protein docking in ZDOCK using an advanced 3D convolution library[J].PLoS ONE, 2011, 6(9): e24657. DOI: 10.1371/journal.pone.0024657.

YAN X, SHI Z F, XU L X, et al . Glutamate impairs mitochondria aerobic respiration capacity and enhances glycolysis in cultured rat astrocytes[J ] . Biomed Environ Sci, 2017, 30(1): 44-51. DOI: 10.3967/bes2017.005.

TORRES M J, KEW K A, RYAN T E, et al . 17β-estradiol directly lowers mitochondrial membranemicroviscosity and improves bioenergetic function in skeletal muscle[J ] . Cell Metab, 2018, 27(1): 167-179.e7. DOI: 10.1016/j.cmet.2017.10.003.

段晨阳, 向鑫明, 匡磊, 等.活化Drp1介导谷胱甘肽代谢调节失血性休克后线粒体功能的研究[J].第三军医大学学报, 2020, 42(1): 50-58. DOI: 10.16016/j.1000-5404.201907139.

DUAN C Y, XIANG X M, KUANG L, et al . Role of activated dynamin-related protein 1-mediated glutathione metabolism in regulation of mitochondrial dysfunction after hemorrhagic shock[J ] . J Third Mil Med Univ, 2020, 42(1): 50-58. DOI:10.16016/j.1000-5404.201907139.

FUHRMANN D C, WITTIG I, HEIDE H, et al . Chronic hypoxia alters mitochondrial composition in human macrophages[J ] .Biochim Biophys Acta, 2013, 1834(12): 2750-2760. DOI: 10.1016/j.bbapap.2013.09.023.

ZAJA I, BAI X W, LIU Y N, et al . Cdk1, PKCδ and calcineurin-mediated Drp1 pathway contributes to mitochondrial fission-induced cardiomyocyte death[J ] . Biochem Biophys Res Commun, 2014, 453(4): 710-721. DOI: 10.1016/j.bbrc.2014.09.144.

WANG W J, WANG Y, LONG J Y, et al . Mitochondrial fission triggered by hyperglycemia is mediated by ROCK1 activation inpodocytes and endothelial cells[J ] . Cell Metab, 2012, 15(2): 186-200. DOI: 10.1016/j.cmet.2012.01.009.

HU C X, HUANG Y, LI L J. Drp1-dependent mitochondrial fission plays critical roles in physiological and pathological progresses in mammals[J].Int J Mol Sci, 2017, 18(1): E144. DOI: 10.3390/ijms18010144.

HORN S R, THOMENIUS M J, JOHNSON E S, et al . Regulation of mitochondrial morphology by APC/CCdh1-mediated control of Drp1 stability[J ] .Mol Biol Cell, 2011, 22(8): 1207-1216. DOI: 10.1091/mbc.E10-07-0567.

PRUDENT J, ZUNINO R, SUGIURA A, et al . MAPL SUMOylation of Drp1 stabilizes an ER/mitochondrial platform required for cell death[J ] . Mol Cell, 2015, 59(6): 941-955. DOI: 10.1016/j.molcel.2015.08.001.

ZHANG J D, FENG J C, MA D, et al .Neuroprotective mitochondrial remodeling by AKAP121/PKA protects HT22 cell from glutamate-induced oxidative stress[J ] . Mol Neurobiol, 2019, 56(8): 5586-5607. DOI: 10.1007/s12035-018-1464-3.

JIN X F, WANG J, GAO K, et al .Dysregulation of INF2-mediated mitochondrial fission in SPOP-mutated prostate cancer[J ] . PLoS Genet, 2017, 13(4): e1006748. DOI: 10.1371/journal.pgen.1006748.

LIU W, ACIN-PEREZ R, GEGHMAN K D, et al . Pink1 regulates the oxidative phosphorylation machinery via mitochondrial fission[J ] .Proc Natl Acad Sci U S A, 2011, 108(31): 12920-12924. DOI: 10.1073/pnas.1107332108..

杨玲, 刘若兰, 牛欢, 等.低表达线粒体蛋白ISCA2对线粒体功能影响的机制研究[J].中国细胞生物学学报, 2018, 40(7): 1120-1129.

YANG L, LIU R L, NIU H, et al . Elucidation of roles of protein ISCA2 in the control of mitochondrial function[J ] . Chin J Cell Biol, 2018, 40(7): 1120-1129.

SUMI C, OKAMOTO A, TANAKA H, et al .Propofol induces a metabolic switch to glycolysis and cell death in a mitochondrial electron transport chain-dependent manner[J ] . PLoS ONE, 2018, 13(2): e0192796. DOI: 10.1371/journal.pone.0192796.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Role of activated dynamin-related protein 1-mediated glutathione metabolism in regulation of mitochondrial dysfunction after hemorrhagic shock

Protective effect of citrus flavone against acute hypoxia in cultured human intestinal smooth muscle cells and the mechanism

Effect of hypoxia on miR-210 expression and mitochondrial function in PC12 cells

Dynamic changes of membrane potentials, cytoplasmic and mitochondrial Ca²⁺ in cultured pulmonary artery smooth muscle cells exposed to hypoxia

Effects of training under hypobaric hypoxia on rat myocardial mitochondrial function

Related Author

Chenyang DUAN

Xinming XIANG

Lei KUANG

Liangming LIU

Tao LI

刘景君

王晓明

沈成凤

Related Institution

State Key Laboratory of Trauma, Burns and Combined Injury, Department of War Wound Shock and Blood Transfusion, Institute of Surgery Research, Army Medical University (Third Military Medical University)

Department of Frigid Zone Medicine， College of High Altitude Military Medicine, School of Nursing， Army Medical University （Third Military Medical University）

Key Laboratory of Extreme Environmental Medicine of Ministry of Education, School of Nursing， Army Medical University （Third Military Medical University）

Department of Clinical Nursing, School of Nursing， Army Medical University （Third Military Medical University）

Institute of Immunology, College of Basic Medical Science，Army Medical University （Third Military Medical University）

⁰