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1. Autor: Velázquez, Diego

8 záznamů v Medvik Filtry

Článek online

Allosteric links between the hydrophilic N-terminus and transmembrane core of human Na+ /H+ antiporter NHA2

Velázquez, Diego
Autor Velázquez, Diego ORCID Laboratory of Membrane Transport, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic
Průša, Vojtěch
Autor Průša, Vojtěch Laboratory of Membrane Transport, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic
Masrati, Gal
Autor Masrati, Gal ORCID Department of Biochemistry and Molecular Biology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv, Israel
Yariv, Elon
Autor Yariv, Elon ORCID Department of Biochemistry and Molecular Biology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv, Israel
Sychrova, Hana
Autor Sychrova, Hana ORCID Laboratory of Membrane Transport, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic
Ben-Tal, Nir
Autor Ben-Tal, Nir ORCID Department of Biochemistry and Molecular Biology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv, Israel
Zimmermannova, Olga
Autor Zimmermannova, Olga ORCID Laboratory of Membrane Transport, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic

Protein science. 2022 ; 31 (12) : e4460. [pub] -

Protein Sci
ISSN 1469-896X
Medvik
Zdroj

The human Na+ /H+ antiporter NHA2 (SLC9B2) transports Na+ or Li+ across the plasma membrane in exchange for protons, and is implicated in various pathologies. It is a 537 amino acids protein with an 82 residues long hydrophilic cytoplasmic N-terminus followed by a transmembrane part comprising 14 transmembrane helices. We optimized the functional expression of HsNHA2 in the plasma membrane of a salt-sensitive Saccharomyces cerevisiae strain and characterized in vivo a set of mutated or truncated versions of HsNHA2 in terms of their substrate specificity, transport activity, localization, and protein stability. We identified a highly conserved proline 246, located in the core of the protein, as being crucial for ion selectivity. The replacement of P246 with serine or threonine resulted in antiporters with altered substrate specificity that were not only highly active at acidic pH 4.0 (like the native antiporter), but also at neutral pH. P246T/S versions also exhibited increased resistance to the HsNHA2-specific inhibitor phloretin. We experimentally proved that a putative salt bridge between E215 and R432 is important for antiporter function, but also structural integrity. Truncations of the first 50-70 residues of the N-terminus doubled the transport activity of HsNHA2, while changes in the charge at positions E47, E56, K57, or K58 decreased the antiporter's transport activity. Thus, the hydrophilic N-terminal part of the protein appears to allosterically auto-inhibit cation transport of HsNHA2. Our data also show this in vivo approach to be useful for a rapid screening of SNP's effect on HsNHA2 activity.

MeSH
lidé MeSH
Na(+)-H(+) antiport * chemie genetika MeSH
protony * MeSH
Saccharomyces cerevisiae genetika metabolismus MeSH
sekvence aminokyselin MeSH
Check Tag
lidé MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Výtvarné umění

Les menines "La familia"

Velázquez, Diego, 1599-1660
Autor Autorita

[Místo vydání není známé] : [nakladatel není známý], [mezi 1900 a 1932?]

1 list ; 46 x 36 cm

Las Meninas', 1656 - Dvorní dámy "Rodina" - Markéta Habsburská (Marguerite-Thérèse d'Autriche) s dvorními dámami, vpravo trpasličí dívka, zachycené při portrétování autorem obrazu.