SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.

Proteins of the conserved Mep-Amt-Rh family, including mammalian Rhesus factors, mediate transmembrane ammonium transport. Ammonium is an important nitrogen source for the biosynthesis of amino acids but is also a metabolic waste product. Its disposal in urine plays a critical role in the regulation...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Aude Deschuyteneer, Mélanie Boeckstaens, Christelle De Mees, Pascale Van Vooren, René Wintjens, Anna Maria Marini
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/159e8141b1444204b6c65186496796da
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:159e8141b1444204b6c65186496796da
record_format	dspace
spelling	oai:doaj.org-article:159e8141b1444204b6c65186496796da2021-11-18T08:59:56ZSNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.1932-620310.1371/journal.pone.0071092https://doaj.org/article/159e8141b1444204b6c65186496796da2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23967154/?tool=EBIhttps://doaj.org/toc/1932-6203Proteins of the conserved Mep-Amt-Rh family, including mammalian Rhesus factors, mediate transmembrane ammonium transport. Ammonium is an important nitrogen source for the biosynthesis of amino acids but is also a metabolic waste product. Its disposal in urine plays a critical role in the regulation of the acid/base homeostasis, especially with an acid diet, a trait of Western countries. Ammonium accumulation above a certain concentration is however pathologic, the cytotoxicity causing fatal cerebral paralysis in acute cases. Alteration in ammonium transport via human Rh proteins could have clinical outcomes. We used a yeast-based expression assay to characterize human Rh variants resulting from non synonymous single nucleotide polymorphisms (nsSNPs) with known or unknown clinical phenotypes and assessed their ammonium transport efficiency, protein level, localization and potential trans-dominant impact. The HsRhAG variants (I61R, F65S) associated to overhydrated hereditary stomatocytosis (OHSt), a disease affecting erythrocytes, proved affected in intrinsic bidirectional ammonium transport. Moreover, this study reveals that the R202C variant of HsRhCG, the orthologue of mouse MmRhcg required for optimal urinary ammonium excretion and blood pH control, shows an impaired inherent ammonium transport activity. Urinary ammonium excretion was RHcg gene-dose dependent in mouse, highlighting MmRhcg as a limiting factor. HsRhCG(R202C) may confer susceptibility to disorders leading to metabolic acidosis for instance. Finally, the analogous R211C mutation in the yeast ScMep2 homologue also impaired intrinsic activity consistent with a conserved functional role of the preserved arginine residue. The yeast expression assay used here constitutes an inexpensive, fast and easy tool to screen nsSNPs reported by high throughput sequencing or individual cases for functional alterations in Rh factors revealing potential causal variants.Aude DeschuyteneerMélanie BoeckstaensChristelle De MeesPascale Van VoorenRené WintjensAnna Maria MariniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71092 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Aude Deschuyteneer Mélanie Boeckstaens Christelle De Mees Pascale Van Vooren René Wintjens Anna Maria Marini SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
description	Proteins of the conserved Mep-Amt-Rh family, including mammalian Rhesus factors, mediate transmembrane ammonium transport. Ammonium is an important nitrogen source for the biosynthesis of amino acids but is also a metabolic waste product. Its disposal in urine plays a critical role in the regulation of the acid/base homeostasis, especially with an acid diet, a trait of Western countries. Ammonium accumulation above a certain concentration is however pathologic, the cytotoxicity causing fatal cerebral paralysis in acute cases. Alteration in ammonium transport via human Rh proteins could have clinical outcomes. We used a yeast-based expression assay to characterize human Rh variants resulting from non synonymous single nucleotide polymorphisms (nsSNPs) with known or unknown clinical phenotypes and assessed their ammonium transport efficiency, protein level, localization and potential trans-dominant impact. The HsRhAG variants (I61R, F65S) associated to overhydrated hereditary stomatocytosis (OHSt), a disease affecting erythrocytes, proved affected in intrinsic bidirectional ammonium transport. Moreover, this study reveals that the R202C variant of HsRhCG, the orthologue of mouse MmRhcg required for optimal urinary ammonium excretion and blood pH control, shows an impaired inherent ammonium transport activity. Urinary ammonium excretion was RHcg gene-dose dependent in mouse, highlighting MmRhcg as a limiting factor. HsRhCG(R202C) may confer susceptibility to disorders leading to metabolic acidosis for instance. Finally, the analogous R211C mutation in the yeast ScMep2 homologue also impaired intrinsic activity consistent with a conserved functional role of the preserved arginine residue. The yeast expression assay used here constitutes an inexpensive, fast and easy tool to screen nsSNPs reported by high throughput sequencing or individual cases for functional alterations in Rh factors revealing potential causal variants.
format	article
author	Aude Deschuyteneer Mélanie Boeckstaens Christelle De Mees Pascale Van Vooren René Wintjens Anna Maria Marini
author_facet	Aude Deschuyteneer Mélanie Boeckstaens Christelle De Mees Pascale Van Vooren René Wintjens Anna Maria Marini
author_sort	Aude Deschuyteneer
title	SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
title_short	SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
title_full	SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
title_fullStr	SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
title_full_unstemmed	SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.
title_sort	snps altering ammonium transport activity of human rhesus factors characterized by a yeast-based functional assay.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/159e8141b1444204b6c65186496796da
work_keys_str_mv	AT audedeschuyteneer snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay AT melanieboeckstaens snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay AT christelledemees snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay AT pascalevanvooren snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay AT renewintjens snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay AT annamariamarini snpsalteringammoniumtransportactivityofhumanrhesusfactorscharacterizedbyayeastbasedfunctionalassay
_version_	1718421007892480000

SNPs altering ammonium transport activity of human Rhesus factors characterized by a yeast-based functional assay.

Ejemplares similares