H2020Индивидуална стипендия2018–2020

NaKStruc · Structural studies of Na,K-ATPase isoforms and mutants

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2018-06-01 → 2020-05-31
Финансиране от ЕС: 200 195 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Протеинът Na,K-ATPase и негови мутации, като например E815K, се анализират, за да се разбере как се променя структурата и работата им. Това е важно, защото тези промени причиняват неврологични заболявания и разбирането им помага при разработването на нови лекарства.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Structural studies of Na,K-ATPase isoforms and mutants

This project focuses on the investigation of disease mutants of Na,K-ATPase. The Na,K-ATPase is a protein found in the cell membrane of virtually every animal cell that maintains the unequal distribution of sodium and potassium between the inside and outside of cells. It is thus vital for numerous functions such as import of other solute, e.g. sugars, neurotransmitters and other ions, cell volume regulation as well as maintaining the electric excitability of neurons and muscle. Mutations of single amino acids have recently been identified as the cause of several neurological diseases. These include Alternating hemiplegia of childhood (AHC), which is characterized by an early onset in infancy, motor impairments such as hemiplegia, i.e. episodic weakness of one half of the body, epileptic seizures and developmental delay. Since there are more than 200 individual mutations causing the disease symptoms vary from milder outcomes to early death. The disease does not only affect individuals, yet depending on severity also their family and caretakers. On of the most severe mutations causing AHC is the E815K mutant, a change of a negatively charged glutamate to a positively charged lysine, which accounts for more than 10% of cases. We aim at studying this mutant and its affect on pump function and the impact on the structure of the protein. We hope that a better understanding of mutations such as E815K will help to pave the way towards cures and that structural insights will lead to drug developments in the near future. We aimed at studying the underlying kinetic mechanism of the E815K-mutation and characterize its impact on the structure of the protein.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Isoform complexes of Na,K-ATPase , an αβ-heterooligomer, have recently been identified as crucial regulators of muscle and neural physiology and were furthermore identified as the primary cause of neurological diseases. Of particular interest are α2β2 & α2β3, which are expressed in muscle and astrocytes (α2β2) and the ciliary body (α2β3). Both exhibit a significantly lower affinity for K-ions and a steeper voltage dependence than the housekeeping α1β1 complex. These features keep α2-isoforms inactive at resting conditions but allow to respond to transient elevations of extracellular K after muscle and neuronal activity. Moreover, both complexes can be selectively inhibited by cardiac steroids, making them promising drug targets for the treatment of heart failure (α2β2) and glaucoma (α2β3). Differences in affinity for K and cardiac steroids were attributed to the β-subunit recently, yet the root cause and molecular details remained elusive. Hence, I propose to investigate the structure of α2-isoforms purified from the yeast Pichia pastoris by x-ray crystallography and cryo electron microscopy. In addition to its physiological and pharmacological role mutations of Na,K-ATPase cause severe neurological diseases, e.g. Alternating Hemiplegia of Childhhod. Disease mutations typically inactivate the Na-pump causing severe motor and cognitive disorder. However, the effects of mutations on the ATPase’s reaction cycle have not been investigated in detail, leaving the cause of inactivation in the dark. Part two of the proposal addresses this gap of knowledge. The two most common mutations E815K and D801N will be expressed in P. pastoris and the effect of mutations on partial reactions of the Na,K-ATPase’s reaction cycle will be investigated and subsequently, structural studies will be carried out. This work will promote our understanding of Na,K-ATPase in health and disease and lay the foundation for the development of new drugs.

Оригинален текст от CORDIS (на английски).

Участници

AARHUS UNIVERSITET · Aarhus CКоординаторДания