ZIB

Treffer pro Seite

Treffer 1 - 2 | 2 Treffer

Sortierung

Digitale Medien

Characterisation of tissue-specific oligosaccharides from rat brain and kidney membrane preparations enriched in Na+,K+-ATPase (1999)

Clark, Richard A.C. ; Kuster, Bernhard ; Benallal, Mounja ; [weitere]

Springer

Glycoconjugate journal 16 (1999), S. 437-456

zur Merkliste hinzufügen auf der Merkliste

Details

ISSN: 1573-4986

Schlagwort(e): glycosylation ; Na+,K+-ATPase ; oligomannose ; polylactosamine ; tissue-specific ; 2-AB, 2-aminobenzamide ; AMOG, adhesion molecule on glia ; ATP, adenosine triphosphate ; BSA, bovine serum albumin ; CAM, calcium/calmodulin-dependent protein kinase ; CNS, central nervous system ; DSA, Datura stramonium agglutinin (lactosamine and sialic acid recognition) ; EDTA, ethylenediaminetetra-acetic acid ; Gal, galactose ; Glc-NAc, N-acetyl-D-glucosamine ; GNA, Galanthus nivalis agglutinin (oligomannosidic recognition) ; GnT, N-acetylglucosaminyltransferase ; GU, glucose unit ; HPLC, high performance liquid chromatography ; MALDI, matrix-assisted laser desorption/ionisation ; (Man)5–9, oligomannose-type glycan series with 5–9 manose residues ; Man1, Manβ1-4GlcNAcβ1-4 GlcNAc ; Man3, trimannose-chitobiose core ; Man3F, trimannose-chitobiose core with fucose on reducing terminal GlcNAc ; MS, mass spectrometry ; NP normal phase ; PBS, phosphate buffered saline ; PNGase F, protein-N-glycosidase F ; PVDF, poly(vinylidene difluoride) ; SDS-PAGE, sodium dodecyl sulphate polyacrylamine gel electrophoresis ; TOF, time-of-flight ; Tris, 2-amino-2-hydroxymethylpropane-1,3 diol ; WAX, weak anion exchange

Quelle: Springer Online Journal Archives 1860-2000

Thema: Chemie und Pharmazie

Notizen: Abstract The organ-specific nature of the glycosylation of Na+,K+-ATPase-enriched preparations from kidney and brain tissues has earlier been indicated by the use of lectin-staining techniques. Na+,K+-ATPase is ubiquitous and abundant, and subject to upregulation during cell-division and in certain pathological conditions. Lectins specific for the different carbohydrates displayed by the Na+,K+-ATPases may, therefore, be useful carriers/mediators in tissue-specific targeting. N-linked oligosaccharides purified from Na+,K+-ATPase-enriched preparations from rat brain and kidney were consequently characterised in detail in this study using weak anion exchange and normal phase HPLC (combined with serial glycosidase digestions) and matrix-assisted laser desorption/ionisation mass spectrometry. The oligomannose series of glycans were most abundant in the brain tissue preparation and this contrasted with the renal-associated oligosaccharides that were dominated by families of tetra-antennary glycans (with/without a core fucose) with up to four lactosaminylglycan residues in either branched or linear formation.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1023/A:1007078511110

Permalink

Bibliothek	Standort	Signatur	Band/Heft/Jahr	Verfügbarkeit

Andere fanden auch interessant ...

Artikel (Nationallizenzen)

Volltext

Digitale Medien

Structural determination of N-linked carbohydrates by matrix-assisted laser desorption/ionization-mass spectrometry following enzymatic release within sodium dodecyl sulphate-polyacrylamide electrophoresis gels: Application to species-specific glycosylation of α1-acid glycoprotein (1998)

Küster, Bernhard ; Hunter, Ann P. ; Wheeler, Susan F. ; [weitere]

Weinheim : Wiley-Blackwell

Electrophoresis 19 (1998), S. 1950-1959

zur Merkliste hinzufügen auf der Merkliste

Details

ISSN: 0173-0835

Schlagwort(e): Carbohydrates ; Matrix assisted laser desorption ; ionization-mass spectrometry ; Sodium dodecyl sulfate - polyacrylamide gel electrophoresis ; Peptide N-glycosidase P ; α1-Acid glycoprotein ; Chemistry ; Biochemistry and Biotechnology

Quelle: Wiley InterScience Backfile Collection 1832-2000

Thema: Biologie , Chemie und Pharmazie

Notizen: This paper describes a sensitive method for analysis of N-linked carbohydrates released enzymatically from within the gel following separation of glycoproteins (50-100 pmols) by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The separated bands containing the glycoproteins were cut from the gel, destained, reduced and alkylated. N-linked glycans were then released by in-gel incubation with peptide N-glycosidase-F (PNGase-F) and extracted with water and acetonitrile. Sialic acid-containing glycans were converted into methyl esters by reaction with methyl iodide, salts and reagents were removed by passage through a mixed-bed column of ion-exchange resins and the glycans were examined by matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry. Structural determination of the released glycans was performed by exoglycosidase digestion. Following glycan release and extraction, the protein could be digested within the gel with trypsin, and the masses of the tryptic peptides could be compared with those generated from a sequence database for protein identification. The method is applied to the analysis of N-linked glycans from α1-acid glycoprotein from man, cow, sheep and dog. Major species-specific differences in glycosylation were found. Thus, although all four species used N-acetyl-neuraminic acid, only cow and sheep additionally used N-glycolyl-neuraminic acid. Biantennary glycans were the predominant carbohydrates in cow, sheep and dog but man produced more triantennary glycans and a substantial amount of tetraantennary sugars. Fucosylation was only found in glycans from man and cow and both cow and sheep glycans were found to have β1-3- and well as β1-4-linked galactose residues in the antennae.

Zusätzliches Material: 4 Ill.