Biomaterial Database

Asn49 is the unique glycosylation site of the trout red blood cell Na+/H+ exchanger. 1998

M Malapert, and B Pellissier, and F Borgese

Laboratoire Jean Maetz, ERS 1253, Département de Biologie Cellulaire et Moléculaire CEA, Villefranche-sur-mer, France.

The Na+/H+ exchanger (NHE) is a plasma membrane transport protein found in a wide range of biological systems. NHE is involved in various functions including pH homeostasis, volume regulation, cell proliferation and transcellular Na+ absorption. This study reports immunodetection results obtained with antibodies generated against the C-terminus of the NHE of trout red blood cells, betaNHE. Immunoblotting of cell membrane preparation reveals that betaNHE is a protein with an apparent molecular mass of 95 kDa. Moreover enzymatic glycosidase treatment demonstrates that the antiporter is an N-glycosylated but not O-glycosylated protein. The primary structure of betaNHE contains three putative N-glycosylation consensus sites (N-X-S/T) at Asn49, Asn338 and Asn378. Expression of betaNHE in PS120 fibroblasts, a cell line which lacks an endogenous Na+/H+ exchange, allows to determine the precise sites of glycosylation. The construction of a site-directed mutated betaNHE antiporter, lacking the first predicted motif, shows that betaNHE possesses an unique glycosylation site located on the first extracellular loop of the exchanger (Asn49). Expression of this deglycosylated antiporter shows that deglycosylation of the protein modifies neither the pH(i) dependency of the antiporter nor its hormonal stimulation.

UI	MeSH Term	Description	Entries
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D003412	Cricetulus	A genus of the family Muridae consisting of eleven species. C. migratorius, the grey or Armenian hamster, and C. griseus, the Chinese hamster, are the two species used in biomedical research.	Hamsters, Armenian,Hamsters, Chinese,Hamsters, Grey,Armenian Hamster,Armenian Hamsters,Chinese Hamster,Chinese Hamsters,Grey Hamster,Grey Hamsters,Hamster, Armenian,Hamster, Chinese,Hamster, Grey
D004912	Erythrocytes	Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.	Blood Cells, Red,Blood Corpuscles, Red,Red Blood Cells,Red Blood Corpuscles,Blood Cell, Red,Blood Corpuscle, Red,Erythrocyte,Red Blood Cell,Red Blood Corpuscle
D006031	Glycosylation	The synthetic chemistry reaction or enzymatic reaction of adding carbohydrate or glycosyl groups. GLYCOSYLTRANSFERASES carry out the enzymatic glycosylation reactions. The spontaneous, non-enzymatic attachment of reducing sugars to free amino groups in proteins, lipids, or nucleic acids is called GLYCATION (see MAILLARD REACTION).	Protein Glycosylation,Glycosylation, Protein
D006224	Cricetinae	A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.	Cricetus,Hamsters,Hamster
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D001216	Asparagine	A non-essential amino acid that is involved in the metabolic control of cell functions in nerve and brain tissue. It is biosynthesized from ASPARTIC ACID and AMMONIA by asparagine synthetase. (From Concise Encyclopedia Biochemistry and Molecular Biology, 3rd ed)	L-Asparagine
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D014337	Trout	Various fish of the family SALMONIDAE, usually smaller than salmon. They are mostly restricted to cool clear freshwater. Some are anadromous. They are highly regarded for their handsome colors, rich well-flavored flesh, and gameness as an angling fish. The genera Salvelinus, Salmo, and ONCORHYNCHUS have been introduced virtually throughout the world.	Chars,Salvelinus,Char
D015153	Blotting, Western	Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.	Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings