The cysteine residue at 424th of pyruvate kinase M2 is crucial for tetramerization and responsiveness to oxidative stress

Masaki, So; Hashimoto, Kozue; Kihara, Daiki; Tsuzuki, Chizuru; Kataoka, Naoyuki; Suzuki, Kenji

doi:10.1016/j.bbrc.2020.03.182

Cited by 15 publications

(6 citation statements)

References 18 publications

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“…This is the first report to identify the oxidative modification of Cys424 induced by H 2 O 2 . In addition to the previous findings [63][64][65], our findings suggest that-as a redox-active cysteine residue-Cys424 might tend to be a thiolate anion. Cys424 is located at the interface of the dimer-dimer interaction of the tetramer PKM2.…”

Section: Discussionsupporting

confidence: 70%

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Comprehensive analyses of the cysteine thiol oxidation of PKM2 reveal the effects of multiple oxidation on cellular oxidative stress response

Irokawa

Numasaki

Kato

et al. 2021

Biochemical Journal

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Redox regulation of proteins via cysteine residue oxidation is involved in the control of various cellular signal pathways. Pyruvate kinase M2 (PKM2), a rate-limiting enzyme in glycolysis, is critical for the metabolic shift from glycolysis to the pentose phosphate pathway under oxidative stress in cancer cell growth. The PKM2 tetramer is required for optimal pyruvate kinase (PK) activity, whereas the inhibition of inter-subunit interaction of PKM2 induced by Cys358 oxidation has reduced PK activity. In the present study, we identified three oxidation-sensitive cysteine residues (Cys358, Cys423 and Cys424) responsible for four oxidation forms via the thiol oxidant diamide and/or hydrogen peroxide (H2O2). Possibly due to obstruction of the dimer-dimer interface, H2O2-induced sulfenylation (-SOH) and diamide-induced modification at Cys424 inhibited tetramer formation and PK activity. Cys423 is responsible for intermolecular disulphide bonds with heterologous proteins via diamide. Additionally, intramolecular polysulphide linkage (–Sn–, n≧3) between Cys358 and an unidentified PKM2 Cys could be induced by diamide. We observed that cells expressing the oxidation-resistant PKM2 (PKM2C358,424A) produced more intracellular reactive oxygen species (ROS) and exhibited greater sensitivity to ROS-generating reagents and ROS-inducible anti-cancer drugs compared to cells expressing wildtype PKM2. These results highlight the possibility that PKM2 inhibition via Cys358 and Cys424 oxidation contributes to eliminating excess ROS and oxidative stress.

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Section: Discussionsupporting

confidence: 70%

“…Thus, sulfenylation and the electrophilic addition of Cys424-SH might induce structural changes on the dimer-dimer interface. Previous studies have revealed that electrophilic compounds such as 4-hydroxy-2-nonenal and 4-oxo-2-nonenal can react with Cys424 and form Michael adducts [63][64][65].…”

Section: Discussionmentioning

confidence: 99%

Comprehensive analyses of the cysteine thiol oxidation of PKM2 reveal the effects of multiple oxidation on cellular oxidative stress response

Irokawa

Numasaki

Kato

et al. 2021

Biochemical Journal

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“…Although the interaction between PKM2 with SERCA2a was via exon 1–8 or 11–12, the same domains as PKM1, we confirmed that PKM1 did not interact with SERCA2a. The reason is that PKM1 and PKM2 differ greatly in their spatial structure, subcellular location, and biochemical functions, due to the difference of only 22 amino acids encoded by exon 10 [ 21 , 48 , 49 ]. PKM2 functions indispensably for the expression of SERCA2a and its exon 10 was the critical structure.…”

Section: Discussionmentioning

confidence: 99%

PKM2 deficiency exacerbates gram-negative sepsis-induced cardiomyopathy via disrupting cardiac calcium homeostasis

Lin

Shen

et al. 2022

Cell Death Discov.

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Sepsis is a life-threatening syndrome with multi-organ dysfunction in critical care medicine. With the occurrence of sepsis-induced cardiomyopathy (SIC), characterized by reduced ventricular contractility, the mortality of sepsis is boosted to 70–90%. Pyruvate kinase M2 (PKM2) functions in a variety of biological processes and diseases other than glycolysis, and has been documented as a cardioprotective factor in several heart diseases. It is currently unknown whether PKM2 influences the development of SIC. Here, we found that PKM2 was upregulated in cardiomyocytes treated with LPS both in vitro and in vivo. Pkm2 inhibition exacerbated the LPS-induced cardiac damage to neonatal rat cardiomyocytes (NRCMs). Furthermore, cardiomyocytes lacking PKM2 aggravated LPS-induced cardiomyopathy, including myocardial damage and impaired contractility, whereas PKM2 overexpression and activation mitigated SIC. Mechanism investigation revealed that PKM2 interacted with sarcoplasmic/endoplasmic reticulum calcium ATPase 2a (SERCA2a), a key regulator of the excitation-contraction coupling, to maintain calcium homeostasis, and PKM2 deficiency exacerbated LPS-induced cardiac systolic dysfunction by impairing SERCA2a expression. In conclusion, these findings highlight that PKM2 plays an essential role in gram-negative sepsis-induced cardiomyopathy, which provides an attractive target for the prevention and treatment of septic cardiomyopathy.

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“…Tumor cells and myocardial cells under pathological conditions both exhibit high expression of M-type pyruvate kinase 2 (PKM2) [5,6] , which catalyzes a rate-limiting step of glycolysis to produce pyruvate and adenosine triphosphate (ATP) from phosphoenolpyruvate and adenosine diphosphate (ADP). PKM2 oscillates among tetrameric, dimeric and monomeric forms [7] . The tetrameric form of PKM2 exerts the enzyme activity of glycolysis [8,9] .…”

Section: Introductionmentioning

confidence: 99%

“…The tetrameric form of PKM2 exerts the enzyme activity of glycolysis [8,9] . However, in tumor cells, elevated oxidative stress could destabilize PKM2 tetramer through redox modi cations on its cysteine residues [5,7,10] . Reduced catalytic activity of PKM2 thus diverts glucose metabolism ux towards the pentose phosphate pathway to generate adequate reducing power against reactive oxygen species (ROS), thus facilitating tumor cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

M-type pyruvate kinase 2 (PKM2) tetramerization alleviates the progression of right ventricle failure by regulating oxidative stress and mitochondrial dynamics

Guo,

Wang,

Qin

et al. 2023

Preprint

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Background: Right ventricle failure (RVF) is a progressive heart disease that has yet to be fully understood at the molecular level. Elevated M-type pyruvate kinase 2 (PKM2) tetramerization alleviates heart failure, but detailed molecular mechanisms remain unclear. Objective: We observed changes in PKM2 tetramerization levels during the progression of right heart failure and in vitro cardiomyocyte hypertrophy and explored the causal relationship between altered PKM2 tetramerization and the imbalance of redox homeostasis in cardiomyocytes, as well as its underlying mechanisms. Ultimately, our goal was to propose rational intervention strategies for the treatment of RVF. Method: We established RVF in Sprague Dawley (SD) rats by intraperitoneal injection of monocrotaline (MCT). The pulmonary artery pressure and right heart function of rats were assessed using transthoracic echocardiography combined with right heart catheterization. TEPP-46 was used both in vivo and in vitro to promote PKM2 tetramerization. Results: We observed that oxidative stress and mitochondrial disorganization were associated with increased apoptosis in the right ventricular tissue of RVF rats. Quantitative proteomics revealed that PKM2 was upregulated during RVF and negatively correlated with the cardiac function. Facilitating PKM2 tetramerization promoted mitochondrial network formation and alleviated oxidative stress and apoptosis during cardiomyocyte hypertrophy. Moreover, enhancing PKM2 tetramer formation improved cardiac mitochondrial morphology, mitigated oxidative stress and alleviated heart failure. Conculsion: Disruption of PKM2 tetramerization contributed to RVF by inducing mitochondrial fragmentation, accumulating ROS, and finally promoted the progression of cardiomyocyte apoptosis. Facilitating PKM2 tetramerization holds potential as a promising therapeutic approach for RVF.

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The cysteine residue at 424th of pyruvate kinase M2 is crucial for tetramerization and responsiveness to oxidative stress

Cited by 15 publications

References 18 publications

Comprehensive analyses of the cysteine thiol oxidation of PKM2 reveal the effects of multiple oxidation on cellular oxidative stress response

Comprehensive analyses of the cysteine thiol oxidation of PKM2 reveal the effects of multiple oxidation on cellular oxidative stress response

PKM2 deficiency exacerbates gram-negative sepsis-induced cardiomyopathy via disrupting cardiac calcium homeostasis

M-type pyruvate kinase 2 (PKM2) tetramerization alleviates the progression of right ventricle failure by regulating oxidative stress and mitochondrial dynamics

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