Crystal Structure of the N112A Mutant of theLight-Driven Sodium Pump KR2

Maliar, N.; Kovalev, K.; Baeken, C.; Balandin, T.; Astashkin, R.; Rulev, M.; Alekseev, A.; Ilyinsky, N.; Rogachev, A.; Chupin, V.; Dolgikh, D.; Kirpichnikov, M.; Gordeliy, V.

Авторы: Maliar Nina, Kovalev Kirill, Baeken Christian, Balandin Taras, Astashkin Roman, Rulev Maksim, Alekseev Alexey, Ilyinsky Nikolay, Rogachev Andrey, Chupin Vladimir, Dolgikh Dmitry, Kirpichnikov Mikhail, Gordeliy Valentin
Журнал: Crystals
Том: 10
Номер: 496
Год издания: 2020
Издательство: MDPI
Местоположение издательства: Basel, Switzerland
Первая страница: 1
Последняя страница: 15
DOI: 10.3390/cryst10060496
Аннотация: The light-driven sodium pump KR2, found in 2013 in the marine bacteria Krokinobactereikastus, serves as a model protein for the studies of the sodium-pumping microbial rhodopsins (NaRs). KR2 possesses a unique NDQ (N112, D116, and Q123) set of the amino acid residues in the functionally relevant positions, named the NDQ motif. The N112 was shown to determine the Na+/H+ selectivity and pumping eciency of the protein. Thus, N112A mutation converts KR2 into an outward proton pump. However, no structural data on the functional conversions of the light-driven sodium pumps are available at the moment. Here we present the crystal structure of the N112A mutant of KR2 in the ground state at the resolution of 2.4 Å. The structure revealed a minor deflection in the central part of the helix C and a double conformation of the L74 residue in themutant. The organization of the retinal Schi base and neighboring water molecules is preserved in the ground state of KR2-N112A. The presented data provide structural insights into the eects of the alterations of the characteristic NDQ motif of NaRs. Our findings also demonstrate that for the rational design of the KR2 variants with modified ion selectivity for optogenetic applications,the structures of the intermediate states of both the protein and its functional variants are required.Keywords: microbial rhodopsin; retinal; ion transport; mutation; NDQ motif; sodium pump;X-ray crystallography
Добавил в систему: Шайтан Константин Вольдемарович

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Crystal Structure of the N112A Mutant of theLight-Driven Sodium Pump KR2статья

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