The yeast Na+/H+ exchanger Nhx1 is an N-linked glycoprotein. Topological implications

Karen M. Wells, Rajini Rao

Research output: Contribution to journalArticle

Abstract

Nhx1, the endosomal Na+/H+ exchanger of Saccharomyces cerevisiae represents the founding member of a newly emerging subfamily of intracellular Na+/H+ exchangers. These proteins share significantly greater sequence homology to one another than to members of the mammalian Na+/H+ exchanger (NHE) family encoding plasma membrane Na+/H+ exchangers. Members of both subtypes are predicted to share a common organization, with an N-terminal transporter domain of transmembrane helices followed by a C-terminal hydrophilic tail. In the present study, we show that Nhx1 is an asparagine-linked glycoprotein and that the sites of glycosylation map to two residues within the C-terminal stretch of the polypeptide. This is the first evidence, to date, for glycosylation of the C-terminal region of any known NHE isoform. Importantly, the mapping of N-linked glycosylation to the C-terminal domain of Nhx1 is indicative of an unexpected membrane topology, particularly with regard to the orientation of the tail region. Although one recent study demonstrated that certain epitopes in the C-terminal domain of NHE3 were accessible from the exoplasmic side of the plasma membrane (Biemesderfer, D., DeGray, B., and Aronson, P. S. (1998) J. Biol. Chem. 273, 12391-12396), numerous other studies implicate a cytosolic disposition for the hydrophilic C-terminal tail of plasma membrane NHE isoforms. Our analysis of the glycosylation of Nhx1 is strongly indicative of residence of at least some portion of the hydrophilic tail domain within the endosomal lumen. These findings imply that the organization of the tail domain may be more complex than previously assumed.

Original languageEnglish (US)
Pages (from-to)3401-3407
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number5
DOIs
StatePublished - Feb 2 2001

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Sodium-Hydrogen Antiporter
Yeast
Glycosylation
Glycoproteins
Yeasts
Cell membranes
Cell Membrane
Protein Isoforms
Asparagine
Sequence Homology
Saccharomyces cerevisiae
Epitopes
Topology
Membranes
Peptides
Proteins

ASJC Scopus subject areas

  • Biochemistry

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The yeast Na+/H+ exchanger Nhx1 is an N-linked glycoprotein. Topological implications. / Wells, Karen M.; Rao, Rajini.

In: Journal of Biological Chemistry, Vol. 276, No. 5, 02.02.2001, p. 3401-3407.

Research output: Contribution to journalArticle

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