Overview on solubilization and lipid reconstitution of Na,K-ATPase: enzyme kinetic and biophysical characterization

Yoneda, Juliana Sakamoto; Sebinelli, Heitor Gobbi; Itri, Rosângela; Ciancaglini, Pietro

doi:10.1007/s12551-020-00616-5

Cited by 17 publications

(17 citation statements)

References 124 publications

(157 reference statements)

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“…In principle, this pleiotropic effect on wildtype α1 could reflect a classic dominant negative effect mediated by heterodimerization of α3 and α1, but we are aware of no evidence that this ever happens. In the older literature, dimerization or tetramerization of α1 was supported by much indirect evidence ( 49 ) and also by coprecipitation of two tagged α1 constructs expressed in insect cells ( 50 ), but dimerization has not been supported by any analysis of structure and may involve scaffolding intermediaries ( 51 ) or adventitious association in detergent ( 52 , 53 ). Instead, we speculate that the impact of L924P α3 on α1 membrane distribution is due to the extent of membrane remodeling, and that it is a UPR effect with potential to be deleterious.…”

Section: Discussionmentioning

confidence: 99%

Misfolding, altered membrane distributions, and the unfolded protein response contribute to pathogenicity differences in Na,K-ATPase ATP1A3 mutations

Arystarkhova

Ozelius

Brashear

et al. 2021

Journal of Biological Chemistry

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Missense mutations in ATP1A3, the α3 isoform of Na,K-ATPase, cause neurological phenotypes that differ greatly in symptoms and severity. A mechanistic basis for differences is lacking, but reduction of activity alone cannot explain them. Isogenic cell lines with endogenous α1 and inducible exogenous α3 were constructed to compare mutation properties. Na,K-ATPase is made in endoplasmic reticulum, but glycan-free catalytic α subunit complexes with glycosylated β subunit in the ER to proceed through Golgi and post-Golgi trafficking. We previously observed classic evidence of protein misfolding in mutations with severe phenotypes: differences in ER retention of endogenous β1 subunit, impaired trafficking of α3, and cytopathology, suggesting that they misfold during biosynthesis. Here we tested two mutations associated with different phenotypes: D923N, which has a median age of onset of hypotonia or dystonia at 3 years, and L924P, with severe infantile epilepsy and profound impairment. Misfolding during biosynthesis in the ER activates the unfolded protein response (UPR), a multi-armed program that enhances protein folding capacity, and if that fails, triggers apoptosis. L924P showed more nascent protein retention in ER than D923N; more ER-associated degradation of α3 (ERAD); larger differences in Na,K-ATPase subunit distributions among subcellular fractions; and greater inactivation of eIF2α, a major defensive step of the UPR. In L924P there was also altered subcellular distribution of endogenous α1 subunit, analogous to a dominant negative effect. Both mutations showed pro-apoptotic sensitization by reduced phosphorylation of BAD. Encouragingly, however, 4-phenylbutyrate (4PBA), a pharmacological corrector, reduced L924P ER retention, increased α3 expression, and restored morphology.

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Section: Discussionmentioning

confidence: 99%

Misfolding, altered membrane distributions, and the unfolded protein response contribute to pathogenicity differences in Na,K-ATPase ATP1A3 mutations

Arystarkhova

Ozelius

Brashear

et al. 2021

Journal of Biological Chemistry

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“…Phosphatidylserine and phosphatidylglycerol as negatively charged lipids promote the bilayer formation of physiological thickness and increased fluidity tends to support optimal Na + , K + -ATPases activity [62]. Na + , K + -ATPases have been involved in cell signaling, interacting with partner proteins [63].…”

Section: Discussionmentioning

confidence: 99%

Surface Properties of Synaptosomes in the Presence of L-Glutamic and Kainic Acids: In Vitro Alteration of the ATPase and Acetylcholinesterase Activities

2021

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Morphologically and functionally identical to brain synapses, the nerve ending particles synaptosomes are biochemically derived membrane structures responsible for the transmission of neural information. Their surface and mechanical properties, measured in vitro, provide useful information about the functional activity of synapses in the brain in vivo. Glutamate and kainic acid are of particular interest because of their role in brain pathology (including causing seizure, migraine, ischemic stroke, aneurysmal subarachnoid hemorrhage, intracerebral hematoma, traumatic brain injury and stroke). The effects of the excitatory neurotransmitter L-glutamic acid and its agonist kainic acid are tested on Na+, K+-ATPase and Mg2+-ATPase activities in synaptic membranes prepared from the cerebral cortex of rat brain tissue. The surface parameters of synaptosome preparations from the cerebral cortex in the presence of L-glutamic and kainic acids are studied by microelectrophoresis for the first time. The studied neurotransmitters promote a significant increase in the electrophoretic mobility and surface electrical charge of synaptosomes at 1–4 h after isolation. The measured decrease in the bending modulus of model bimolecular membranes composed of monounsaturated lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine provides evidence for softer membranes in the presence of L-glutamate. Kainic acid does not affect membrane mechanical stability even at ten-fold higher concentrations. Both the L-glutamic and kainic acids reduce acetylcholinesterase activity and deviation from the normal functions of neurotransmission in synapses is presumed. The presented results regarding the modulation of the enzyme activity of synaptic membranes and surface properties of synaptosomes are expected by biochemical and biophysical studies to contribute to the elucidation of the molecular mechanisms of neurotransmitters/agonists’ action on membranes.

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“…The Na + /K + -ATPase (NKA), or the sodium-potassium pump, is a ubiquitously expressed plasma membrane protein belonging to the family of P-type ATPases. This group of enzymes use the energy from ATP hydrolysis to pump ions over the plasma membrane against their concentration gradient by autophosphorylation of key aspartate residue [ 73 , 74 ]. NKA pumps 3 Na + out of the cell and 2K + into the cell per single molecule of ATP hydrolyzed, thus maintaining a higher concentration of sodium ions outside and a higher level of potassium ions inside the cell.…”

Section: Na + /K + -Atpasementioning

confidence: 99%

Start Me Up: How Can Surrounding Gangliosides Affect Sodium-Potassium ATPase Activity and Steer towards Pathological Ion Imbalance in Neurons?

et al. 2022

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Gangliosides, amphiphilic glycosphingolipids, tend to associate laterally with other membrane constituents and undergo extensive interactions with membrane proteins in cis or trans configurations. Studies of human diseases resulting from mutations in the ganglioside biosynthesis pathway and research on transgenic mice with the same mutations implicate gangliosides in the pathogenesis of epilepsy. Gangliosides are reported to affect the activity of the Na+/K+-ATPase, the ubiquitously expressed plasma membrane pump responsible for the stabilization of the resting membrane potential by hyperpolarization, firing up the action potential and ion homeostasis. Impaired Na+/K+-ATPase activity has also been hypothesized to cause seizures by several mechanisms. In this review we present different epileptic phenotypes that are caused by impaired activity of Na+/K+-ATPase or changed membrane ganglioside composition. We further discuss how gangliosides may influence Na+/K+-ATPase activity by acting as lipid sorting machinery providing the optimal stage for Na+/K+-ATPase function. By establishing a distinct lipid environment, together with other membrane lipids, gangliosides possibly modulate Na+/K+-ATPase activity and aid in “starting up” and “turning off” this vital pump. Therefore, structural changes of neuronal membranes caused by altered ganglioside composition can be a contributing factor leading to aberrant Na+/K+-ATPase activity and ion imbalance priming neurons for pathological firing.

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Overview on solubilization and lipid reconstitution of Na,K-ATPase: enzyme kinetic and biophysical characterization

Cited by 17 publications

References 124 publications

Misfolding, altered membrane distributions, and the unfolded protein response contribute to pathogenicity differences in Na,K-ATPase ATP1A3 mutations

Misfolding, altered membrane distributions, and the unfolded protein response contribute to pathogenicity differences in Na,K-ATPase ATP1A3 mutations

Surface Properties of Synaptosomes in the Presence of L-Glutamic and Kainic Acids: In Vitro Alteration of the ATPase and Acetylcholinesterase Activities

Start Me Up: How Can Surrounding Gangliosides Affect Sodium-Potassium ATPase Activity and Steer towards Pathological Ion Imbalance in Neurons?

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