Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Keratan sulfate disaccharide composition determined by FACE analysis of keratanase II and endo-beta-galactosidase digestion products
Autore:
Plaas, AHK; West, LA; Midura, RJ;
Indirizzi:
Shriners Hosp Children, Tampa, FL 33612 USA Shriners Hosp Children Tampa FL USA 33612 p Children, Tampa, FL 33612 USA Univ S Florida, Coll Med, Dept Biochem & Mol Biol, Tampa, FL 33620 USA Univ S Florida Tampa FL USA 33620 Biochem & Mol Biol, Tampa, FL 33620 USA Cleveland Clin Fdn, Lerner Res Inst Florida, Cleveland, OH USA Cleveland Clin Fdn Cleveland OH USA Res Inst Florida, Cleveland, OH USA
Titolo Testata:
GLYCOBIOLOGY
fascicolo: 10, volume: 11, anno: 2001,
pagine: 779 - 790
SICI:
0959-6658(200110)11:10<779:KSDCDB>2.0.ZU;2-M
Fonte:
ISI
Lingua:
ENG
Soggetto:
BOVINE ARTICULAR-CARTILAGE; ASSISTED CARBOHYDRATE ELECTROPHORESIS; NONREDUCING CHAIN TERMINI; N-ACETYLNEURAMINIC ACID; MONOCLONAL-ANTIBODIES; ESCHERICHIA-FREUNDII; LARGER OLIGOSACCHARIDES; BACTEROIDES-FRAGILIS; CHONDROITIN SULFATE; FUCOSE RESIDUES;
Keywords:
glycosaminoglycans; keratan sulfate; keratanases; proteoglycans; sulfation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
63
Recensione:
Indirizzi per estratti:
Indirizzo: Plaas, AHK Shriners Hosp Children, 12502 N Pine Dr, Tampa, FL 33612 USA Shriners Hosp Children 12502 N Pine Dr Tampa FL USA 33612 2 USA
Citazione:
A.H.K. Plaas et al., "Keratan sulfate disaccharide composition determined by FACE analysis of keratanase II and endo-beta-galactosidase digestion products", GLYCOBIOLOG, 11(10), 2001, pp. 779-790

Abstract

Many tissues contain glycoproteins and proteoglycans, which are substituted with N-or O-linked keratan sulfate, a glycosaminoglycan in which the lactosamine (-gal beta1,4glcNAc-) disaccharide backbone is variably modified bysulfation, fucosylation, and sialylation. We report here a rapid, sensitive, and quantitative procedure for obtaining a complete disaccharide compositional analyses for keratan sulfates after FACE separation of products generated by hydrolysis of the glycosaminoglycans with B. fragillis keratanase II and E. freundii endo-p-galactosidase. Seven digestion end products are separable in a single electrophoretic step using Monosaccharide(TM) composition gels. These are: the unsulfated disaccharide, glcNAc beta1,3gal, the fucosylated trisaccharide, gal beta1,2[fuc alpha1,3]glcNAc6S, the mono- and disulfated disaccharides, gal beta1,4glcNAc6S or gal6S beta1,4glcNAc6S from the chain interior, and the sialylated mono- and disulfated trisaccharides neuA alpha2,3gal beta1, 4glcNAc6S or neuA alpha2,3gal-6S beta1,4glcNAc6S from the nonreducing terminus. FACE analyses also revealed the presence of a contaminant P-galactosidase activity in keratanase II enzyme preparations which cleaves the disaccharide, gal beta1,4glcNAc6S to its constituent monosaccharides, gal and glcNAc6S. It was particularly prominent at enzyme concentrations > 2 mU per nmole substrate glcNH(2) or after prolonged digestion times (>12 h), and was not inhibitable by thiogalactosides or N-acetyl-lactosamine. As these monosaccharide products would not be detectable using thecommonly described analytical methods for KS hydrolase products, such as H-1-NMR and RPLC analyses, our data illustrate that the FACE procedure represents an improved approach for accurate compositional microanalyses of corneal and skeletal keratan sulfates, especially applicable to experimentationinvolving small amounts (1-2 mug) of this glycosaminoglyean.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/06/20 alle ore 09:44:03