Mitochondrial Calcium Handling in Physiology and Disease

Granatiero, Veronica; Stefani, Diego De; Meneghesso, Gaudenzio

doi:10.1007/978-3-319-55330-6_2

Cited by 66 publications

(55 citation statements)

References 146 publications

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“…Calcium is a key modulator in a range of immunological, metabolic and developmental functions [199][200][201][202]. Calcium signalling is linked to a wide range of physiological and pathological processes [203][204][205], is a key driver for post-translational deimination [206][207][208] and plays an important role in the regulation of EVs [11,48]. In cattle, calcium is linked to many immunological pathways including inflammatory diseases [209], virus replication [210], parasitic infection [211], wound healing [212], reproduction [213,214] and adrenal function [215].…”

Section: Discussionmentioning

confidence: 99%

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

Criscitiello

Kraev

Lange

2020

IJMS

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The bovine immune system is known for its unusual traits relating to immunoglobulin and antiviral responses. Peptidylarginine deiminases (PADs) are phylogenetically conserved enzymes that cause post-translational deimination, contributing to protein moonlighting in health and disease. PADs also regulate extracellular vesicle (EV) release, forming a critical part of cellular communication. As PAD-mediated mechanisms in bovine immunology and physiology remain to be investigated, this study profiled deimination signatures in serum and serum-EVs in Bos taurus. Bos EVs were poly-dispersed in a 70-500 nm size range and showed differences in deiminated protein cargo, compared with whole sera. Key immune, metabolic and gene regulatory proteins were identified to be post-translationally deiminated with some overlapping hits in sera and EVs (e.g., immunoglobulins), while some were unique to either serum or serum-EVs (e.g., histones). Protein-protein interaction network analysis of deiminated proteins revealed KEGG pathways common for serum and serum-EVs, including complement and coagulation cascades, viral infection (enveloped viruses), viral myocarditis, bacterial and parasitic infections, autoimmune disease, immunodeficiency intestinal IgA production, B-cell receptor signalling, natural killer cell mediated cytotoxicity, platelet activation and hematopoiesis, alongside metabolic pathways including ferroptosis, vitamin digestion and absorption, cholesterol metabolism and mineral absorption. KEGG pathways specific to EVs related to HIF-1 signalling, oestrogen signalling and biosynthesis of amino acids. KEGG pathways specific for serum only, related to Epstein-Barr virus infection, transcription mis-regulation in cancer, bladder cancer, Rap1 signalling pathway, calcium signalling pathway and ECM-receptor interaction. This indicates differences in physiological and pathological pathways for deiminated proteins in serum-EVs, compared with serum. Our findings may shed light on pathways underlying a number of pathological and anti-pathogenic (viral, bacterial, parasitic) pathways, with putative translatable value to human pathologies, zoonotic diseases and development of therapies for infections, including anti-viral therapies.

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Section: Discussionmentioning

confidence: 99%

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

Criscitiello

Kraev

Lange

2020

IJMS

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“…The uptake of Ca 2+ into mitochondria has long been established as a key regulator of an array of cellular homeostatic processes as diverse as bioenergetics and cell death [1,2]. A series of seminal discoveries have recently elucidated the identity of the components that make up the mitochondrial Ca 2+ uniporter complex.…”

Section: Introductionmentioning

confidence: 99%

A comprehensive genetic characterisation of the mitochondrial Ca²⁺ uniporter in Drosophila

Tufi¹,

Gleeson²,

Stockum³

et al. 2018

Preprint

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Mitochondrial Ca 2+ uptake is an important mediator of metabolism and cell death. Identification of components of the highly conserved mitochondrial Ca 2+ uniporter has opened it up to genetic analysis in model organisms. Here we report a comprehensive genetic characterisation of the known uniporter components conserved in Drosophila. While loss of MCU or EMRE abolishes fast mitochondrial Ca 2+ uptake, this results in surprisingly mild phenotypes. In contrast, loss of the regulatory gatekeeper component MICU1 has a much more severe phenotype, being developmental lethal, consistent with unregulated Ca 2+ uptake. Mutants for MICU3 are viable with mild neurological phenotypes. Genetic interaction studies reveal that MICU1 and MICU3 are not functionally interchangeable. More surprisingly, loss of MCU or EMRE does not suppress MICU1 mutant lethality, suggesting that the lethality results from MCU-independent functions. This study helps shed light on the physiological requirements of the mitochondrial Ca 2+ uniporter, and provides a suite of tools to interrogate their interplay in homeostasis and disease conditions. B. DNA analysis of MCU 1 . Primer sequences are detailed in the 'Materials and Methods' section.The control genotype yielded a ~2.5 kb band, compared to ~900 bp for MCU 1 homozygotes. C. Western blot analysis of MCU 1 . Immunoblots were probed with the indicated antibodies. Asterisk denotes a non-specific band. Mitochondrial ATP5A is used as a loading control. D.Representative traces of Ca 2+ uptake in mitochondria isolated from adult flies of the indicated genotypes after addition of 45 μM CaCl2. Extramitochondrial Ca 2+ was measured by Calcium Green-5N fluorescence. Ca 2+ was released from mitochondria by addition of 1 μM FCCP. a.u.: arbitrary units. The control genotype is w 1118 . Addition of the MCU inhibitor Ruthenium Red (RuR; 2 μM) blocks mitochondrial Ca 2+ uptake, which is mirrored by MCU 1 . Mitochondrial Ca 2+ uptake is restored by transgenic re-expression of MCU. E. Relative uptake kinetics were determined through linear fits of the initial phase of Ca 2+ uptake and normalized to the wild type control (mean ± SEM, n = 3). Figure 2. MCU 1 shortens lifespan without impacting organismal phenotypes despite respiratory defects. A. The percentage of adult flies eclosing as homozygous MCU 1 mutants versus balanced heterozygotes, together with the expected Mendelian ratio in the offspring (n > 700). B. Lifespan curves of MCU 1 male flies compared with control (w 1118 ) and transgenic rescue (MCU 1 + MCU). Statistical analysis: Mantel-Cox log-rank test (n ≧ 74). C. Climbing assay of control (da/+) and MCU 1 flies, 2 and 20 days post-eclosion. Significance measured by Kruskal-Wallis test with Dunn's post-hoc correction for multiple comparisons (mean ± 95% confidence interval; n > 50; ns, non-significant). D. Relative ATP levels from control and MCU 1 flies. Statistical analysis: unpaired t-test (mean ± SD; n 2-3; **** P < 0.0001, ns, non-significant). E, F. Oxygen consumption rate (OCR) of control and MC...

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“…Ca 2+ binding to the lipid headgroup region severely alters properties of anionic bilayers (48). More relevant to mitochondrial membranes, Ca 2+ interacts strongly with CL, altering bilayer properties (e.g., demixing, phase behavior, and headgroup conformation) (19); Ca 2+ can disrupt protein-lipid interactions, causing effects such as respiratory complex disintegration (54); and mitochondrial Ca 2+ dyshomeostasis underpins a wide variety of mitochondrial diseases (55). In this study, we have directly shown that the binding of SS peptides reduces the effects of Ca 2+ on mitochondrial surface electrostatics ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Molecular Mechanism of Action of Mitochondrial Therapeutic SS-31 (Elamipretide): Membrane Interactions and Effects on Surface Electrostatics

Mitchell

Tamucci

et al. 2019

Preprint

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SignificanceSzeto-Schiller (SS) peptides are among the most promising therapeutic compounds for mitochondrial dysfunction. However, the molecular target(s) and the mechanism of action of SS peptides are poorly understood. In this study, we evaluate the interaction of the lead compound SS-31 (Elamipretide) with mitochondrial and synthetic model membranes using a host of biophysical techniques. Our results show that SS-31 membrane interaction is driven largely by the negative surface charge of mitochondrial membranes and that SS-31 alters lipid bilayer properties, most notably electrostatics at the membrane interface. This work supports a mechanism in which SS peptides act on a key physical property of mitochondrial membranes rather than with a specific protein complex, consistent with the exceptionally broad therapeutic efficacy of these compounds. AbstractMitochondrial dysfunction includes heritable diseases, acquired pathologies, and age-related declines in health. Szeto-Schiller (SS) peptides comprise a class of amphipathic tetrapeptides that have demonstrated efficacy in treating a wide array of mitochondrial disorders, and are believed to target mitochondrial membranes due to their enrichment in the anionic phospholipid cardiolipin (CL). However, little is known regarding how SS peptides interact with or alter the physical properties of lipid bilayers. In this study, we have analyzed the interactions of the lead compound SS-31 (Elamipretide) with model and mitochondrial membranes using biophysical and computational approaches. Our results show that this polybasic peptide partitions into the membrane interfacial region with affinity and binding density that are directly related to surface charge. SS-31 binding does not destabilize lamellar bilayers even at the highest binding concentrations; however, it does cause saturable alterations in lipid packing. Most notably, SS-31 modulates the surface electrostatic properties of model and mitochondrial membranes, which could play a significant role in the mitoprotective properties of this compound. As a proof of concept, we show that SS-31 alters ion distribution at the membrane interface with implications for maintaining mitochondrial membranes subject to divalent cation (calcium) stress. Taken together, these results support a mechanism of action in which SS peptides interact with lipid bilayers and alter the biophysical (primarily electrostatic) properties of mitochondrial membranes as their primary mechanism of action. Understanding this molecular mechanism is key to the development of future compound variants with enhanced efficacy.

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Mitochondrial Calcium Handling in Physiology and Disease

Cited by 66 publications

References 146 publications

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

A comprehensive genetic characterisation of the mitochondrial Ca²⁺ uniporter in Drosophila

Molecular Mechanism of Action of Mitochondrial Therapeutic SS-31 (Elamipretide): Membrane Interactions and Effects on Surface Electrostatics

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Mitochondrial Calcium Handling in Physiology and Disease

Cited by 66 publications

References 146 publications

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

A comprehensive genetic characterisation of the mitochondrial Ca2+ uniporter in Drosophila

Molecular Mechanism of Action of Mitochondrial Therapeutic SS-31 (Elamipretide): Membrane Interactions and Effects on Surface Electrostatics

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A comprehensive genetic characterisation of the mitochondrial Ca²⁺ uniporter in Drosophila