scholarly journals Identification of Phosphorylated Calpain 3 in Rat Brain Mitochondria under mPTP Opening

2021 ◽  
Vol 22 (19) ◽  
pp. 10613
Author(s):  
Yulia Baburuna ◽  
Linda Sotnikova ◽  
Olga Krestinina
Keyword(s):  
Mass Spectrometry ◽  
Rat Brain ◽  
Mitochondrial Permeability Transition ◽  
Tyrosine Phosphatase ◽  
Permeability Transition ◽  
Brain Mitochondria ◽  
Calpain Inhibitor ◽  
Rat Brain Mitochondria ◽  
Calpain 3 ◽  
Synaptic Mitochondria

The protein phosphorylation of the membrane-bound mitochondrial proteins has become of interest from the point of view of its regulatory role of the function of the respiratory chain, opening of the mitochondrial permeability transition pore (mPTP), and initiation of apoptosis. Earlier, we noticed that upon phosphorylation of proteins in some proteins, the degree of their phosphorylation increases with the opening of mPTP. Two isoforms of myelin basic protein and cyclic nucleotide phosphodiesterase were identified in rat brain non-synaptic mitochondria and it was concluded that they are involved in mPTP regulation. In the present study, using the mass spectrometry method, the phosphorylated protein was identified as Calpain 3 in rat brain non-synaptic mitochondria. In the present study, the phosphoprotein Calpain-3 (p94) (CAPN3) was identified in the rat brain mitochondria as a phosphorylated truncated form of p60–62 kDa by two-dimensional electrophoresis and mass spectrometry. We showed that the calpain inhibitor, calpeptin, was able to suppress the Ca2+ efflux from mitochondria, preventing the opening of mPTP. It was found that phosphorylated truncated CALP3 with a molecular weight of 60–62 contains p-Tyr, which indicates the possible involvement of protein tyrosine phosphatase in this process.

Calcium-induced permeability transition in rat brain mitochondria is promoted by carbenoxolone through targeting connexin43

2011 ◽  
Vol 300 (3) ◽  
pp. C707-C720 ◽  
Author(s):  
Tamara Azarashvili ◽  
Yulia Baburina ◽  
Dmitry Grachev ◽  
Olga Krestinina ◽  
Yuri Evtodienko ◽  
...  
Keyword(s):  
Rat Brain ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Brain Mitochondria ◽  
Molecular Target ◽  
Threshold Concentration ◽  
Rat Liver Mitochondria ◽  
Rat Brain Mitochondria ◽  
Synaptic Mitochondria ◽  
Ptp Opening

Carbenoxolone (Cbx), a substance from medicinal licorice, is used for antiinflammatory treatments. We investigated the mechanism of action of Cbx on Ca2+-induced permeability transition pore (PTP) opening in synaptic and nonsynaptic rat brain mitochondria (RBM), as well as in rat liver mitochondria (RLM), in an attempt to identify the molecular target of Cbx in mitochondria. Exposure to threshold Ca2+ load induced PTP opening, as seen by sudden Ca2+ efflux from the mitochondrial matrix and membrane potential collapse. In synaptic RBM, Cbx (1 μM) facilitated the Ca2+-induced, cyclosporine A-sensitive PTP opening, while in nonsynaptic mitochondria the Cbx threshold concentration was higher. A well-known molecular target of Cbx is the connexin (Cx) family, gap junction proteins. Moreover, Cx43 was previously found in heart mitochondria and attributed to the preconditioning mechanism of protection. Thus, we hypothesized that Cx43 might be a target for Cbx in brain mitochondria. For the first time, we detected Cx43 by Western blot in RBM, but Cx43 was absent in RLM. Interestingly, two anti-Cx43 antibodies, directed against amino acids 252 to 270 of rat Cx43, abolished the Cbx-induced enhancement of PTP opening in total RBM and in synaptic mitochondria, but not in RLM. In total RBM and in synaptic mitochondria, PTP caused dephosphorylation of Cx43 at serine 368. The phosphorylation level of serine 368 was decreased at threshold calcium concentration and additionally in the combined presence of Cbx in synaptic mitochondria. In conclusion, active mitochondrial Cx43 appears to counteract the Ca2+-induced PTP opening and thus might inhibit the PTP-ensuing mitochondrial demise and cell death. Consequently, we suggest that activity of Cx43 in brain mitochondria represents a novel molecular target for protection.

Mass spectrometry-based survey of age-associated protein carbonylation in rat brain mitochondria

2007 ◽  
Vol 42 (12) ◽  
pp. 1583-1589 ◽  
Author(s):  
Laszlo Prokai ◽  
Liang-Jun Yan ◽  
José L. Vera-Serrano ◽  
Stanley M. Stevens ◽  
Michael J. Forster
Keyword(s):  
Mass Spectrometry ◽  
Rat Brain ◽  
Brain Mitochondria ◽  
Protein Carbonylation ◽  
Rat Brain Mitochondria

Agmatine prevents the Ca2+-dependent induction of permeability transition in rat brain mitochondria

Amino Acids ◽  
2009 ◽  
Vol 38 (2) ◽  
pp. 431-437 ◽  
Author(s):  
V. Battaglia ◽  
S. Grancara ◽  
J. Satriano ◽  
S. Saccoccio ◽  
E. Agostinelli ◽  
...  
Keyword(s):  
Rat Brain ◽  
Permeability Transition ◽  
Brain Mitochondria ◽  
Rat Brain Mitochondria

Ca2+-dependent permeability transition regulation in rat brain mitochondria by 2′,3′-cyclic nucleotides and 2′,3′-cyclic nucleotide 3′-phosphodiesterase

2009 ◽  
Vol 296 (6) ◽  
pp. C1428-C1439 ◽  
Author(s):  
Tamara Azarashvili ◽  
Olga Krestinina ◽  
Anastasia Galvita ◽  
Dmitry Grachev ◽  
Yulia Baburina ◽  
...  
Keyword(s):  
Rat Brain ◽  
Cyclic Nucleotides ◽  
Cyclic Nucleotide ◽  
Permeability Transition ◽  
Liver Mitochondria ◽  
Brain Mitochondria ◽  
Rat Liver Mitochondria ◽  
Marker Enzyme ◽  
Rat Brain Mitochondria ◽  
Specific Enzymatic Activity

Recent evidence indicates that 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNP), a marker enzyme of myelin and oligodendrocytes, is also present in neural and nonneural mitochondria. However, its role in mitochondria is still completely unclear. We found CNP in rat brain mitochondria and studied the effects of CNP substrates, 2′,3′-cyclic nucleotides, on functional parameters of rat brain mitochondria. 2′,3′-cAMP and 2′,3′-cNADP stimulated Ca2+ overload-induced Ca2+ release from mitochondrial matrix. This Ca2+ release under threshold Ca2+ load correlated with membrane potential dissipation and mitochondrial swelling. The effects of 2′,3′-cyclic nucleotides were suppressed by cyclosporin A, a potent inhibitor of permeability transition (PT). PT development is a key stage in initiation of apoptotic mitochondria-induced cell death. 2′,3′-cAMP effects were observed on the functions of rat brain mitochondria only when PT was developed. This demonstrates involvement of 2′,3′-cAMP in PT regulation in rat brain mitochondria. We also discovered that, under PT development, the specific enzymatic activity of CNP was reduced. Thus we hypothesize that suppression of CNP activity under threshold Ca2+ load leads to elevation of 2′,3′-cAMP levels that, in turn, promote PT development in rat brain mitochondria. Similar effects of 2′,3′-cyclic nucleotides were observed in rat liver mitochondria. Involvement of CNP in PT regulation was confirmed in experiments using mitochondria from CNP-knockdown oligodendrocytes (OLN93 cells). CNP reduction in these mitochondria correlated with lowering the threshold for Ca2+ overload-induced Ca2+ release. Thus our results reveal a new function for CNP and 2′,3′-cAMP in mitochondria, being a regulator/promotor of mitochondrial PT.

scholarly journals Possible Involvement of 2′,3′-Cyclic Nucleotide-3′-Phosphodiesterase in the Protein Phosphorylation-Mediated Regulation of the Permeability Transition Pore

2018 ◽  
Vol 19 (11) ◽  
pp. 3499 ◽  
Author(s):  
Yulia Baburina ◽  
Irina Odinokova ◽  
Tamara Azarashvili ◽  
Vladimir Akatov ◽  
Linda Sotnikova ◽  
...  
Keyword(s):  
Protein Phosphorylation ◽  
Rat Brain ◽  
Protein Kinases ◽  
Cyclic Nucleotide ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Brain Mitochondria ◽  
Calmodulin Antagonist ◽  
Rat Brain Mitochondria ◽  
Mptp Opening

Calcium as a secondary messenger regulates the phosphorylation of several membrane-bound proteins in brain and liver mitochondria. Regulation of the activity of different protein kinases and phosphatases by Ca2+ occurs through its binding with calmodulin. The protein phosphorylation is strongly dependent on the Ca2+-induced mitochondrial permeability transition pore (mPTP) opening. 2′,3′-Cyclic nucleotide-3′-phosphodiesterase (CNPase) was phosphorylated by protein kinases A and C. CNPase and melatonin (MEL) might interact with calmodulin. The effects of the calmodulin antagonist calmidazolium and the inhibitor of protein kinase A H89 on mPTP opening in rat brain mitochondria of male Wistar rats were investigated. In addition, the role of CNPase, serine/threonine kinases, and MEL in the mPTP opening was examined. The anti-CNPase antibody added to rat brain mitochondria (RBM) reduced the content of CNPase in mitochondria. The threshold [Ca2+] decreased, and mitochondrial swelling was accelerated in the presence of the anti-CNPase antibody. H89 enhanced the effect of anti-CNPase antibody and accelerated the swelling of mitochondria, while CmZ abolished the effect of anti-CNPase antibody under mPTP opening. The levels of phospho-Akt and phospho-GSK3β increased, while the MEL content did not change. It can be assumed that CNPase may be involved in the regulation of these kinases, which in turn plays an important role in mPTP functioning.

scholarly journals Calcium‐induced permeability transition in rat brain mitochondria is promoted by carbenoxolone through targeting connexin43

2010 ◽  
Vol 24 (S1) ◽  
Author(s):  
Georg Reiser ◽  
Tamara Azarashvili ◽  
Yulia Baburina ◽  
Dmitry Grachev ◽  
Olga Krestinina
Keyword(s):  
Rat Brain ◽  
Permeability Transition ◽  
Brain Mitochondria ◽  
Rat Brain Mitochondria
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