Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
44
|
| pubmed:dateCreated |
1994-12-6
|
| pubmed:abstractText |
A polyclonal antibody (Ab597) was produced in rabbit against a fusion protein of glutathione-S-transferase and the last 87 amino acids of the Na+/H+ exchanger isoform, NHE2. By Western blotting, Ab597 recognized proteins of 75 and 85 kDa in PS120/NHE2 membranes (PS120 cells stably transfected with NHE2), and this antibody did not cross-react with NHE1 and NHE3. When Ab597 was used to immunocytochemically stain PS120/NHE2 cells, permeabilization of the cells was required for staining, confirming the putative membrane topology of NHE2 that the C-terminus is cytoplasmic. NHE1 is N-glycosylated. NHE2 was predicted to be N-glycosylated as it contains one potential N-linked glycosylation site (N350VS), which is conserved among NHE1, NHE3, and NHE4. However, NHE2 was resistant to peptide:N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) digestion, suggesting that NHE2 is not N-glycosylated. In contrast, neuraminidase shifted the mobility of the 85 kDa NHE2 protein in PS120/NHE2 membranes into an 81 kDa band, and O-glycanase further shifted the mobility of the neuraminidase-treated 81 kDa protein to 75 kDa. Incubation of PS120/NHE2 cells with benzyl N-acetyl-alpha-D-galactosaminide (Bz alpha GalNAc), an O-glycosylation inhibitor, decreased the size of the 85 kDa protein to 81 kDa. This treatment had no effect on the initial rate of Na+/H+ exchange of PS120/NHE2 cells. The 75 kDa protein was not affected by the glycosidase treatment of PS120/NHE2 membranes or the Bz alpha GalNAc treatment of PS120/NHE2 cells.(ABSTRACT TRUNCATED AT 250 WORDS)
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Acetylgalactosamine,
http://linkedlifedata.com/resource/pubmed/chemical/Benzyl Compounds,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Glutathione Transferase,
http://linkedlifedata.com/resource/pubmed/chemical/Immune Sera,
http://linkedlifedata.com/resource/pubmed/chemical/Sialoglycoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Sodium-Hydrogen Antiporter,
http://linkedlifedata.com/resource/pubmed/chemical/benzyl-alpha-N-acetylgalactosamine
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0006-2960
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
8
|
| pubmed:volume |
33
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
12954-61
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:7524659-Acetylgalactosamine,
pubmed-meshheading:7524659-Animals,
pubmed-meshheading:7524659-Benzyl Compounds,
pubmed-meshheading:7524659-Blotting, Western,
pubmed-meshheading:7524659-Cell Line,
pubmed-meshheading:7524659-Cell Membrane Permeability,
pubmed-meshheading:7524659-Cells, Cultured,
pubmed-meshheading:7524659-Cross Reactions,
pubmed-meshheading:7524659-DNA, Complementary,
pubmed-meshheading:7524659-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:7524659-Enzyme Induction,
pubmed-meshheading:7524659-Fibroblasts,
pubmed-meshheading:7524659-Glutathione Transferase,
pubmed-meshheading:7524659-Glycosylation,
pubmed-meshheading:7524659-Immune Sera,
pubmed-meshheading:7524659-Immunohistochemistry,
pubmed-meshheading:7524659-Molecular Weight,
pubmed-meshheading:7524659-Rabbits,
pubmed-meshheading:7524659-Sialoglycoproteins,
pubmed-meshheading:7524659-Skin,
pubmed-meshheading:7524659-Sodium-Hydrogen Antiporter,
pubmed-meshheading:7524659-Staining and Labeling,
pubmed-meshheading:7524659-Transfection
|
| pubmed:year |
1994
|
| pubmed:articleTitle |
Na+/H+ exchanger-2 is an O-linked but not an N-linked sialoglycoprotein.
|
| pubmed:affiliation |
Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|