S1.33 Partial purification and characterization of an endo-?-N-acetylgalactosaminidase from the culture medium ofStreptomyces Sp. OH-11242

Ishii-Karakasa, Ikuko; Iwase, Hitoo; Hotta, Kyoko

doi:10.1007/bf01209834

Cited by 5 publications

(12 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using an assay method established for the measurement of the activity of endoGalNAc-ase-S [2], this enzyme can be isolated from the glycosidase mixture contained in the culture medium. Moreover, this method can also distinguish endo-GalNAc-ase-S from endoGalNAc-ase, an enzyme which has been known to release only Gal/ll-3GalNAc.…”

Section: Resultsmentioning

confidence: 99%

“…One unit (U) of the enzyme activity is defined as that amount of enzyme that catalyzes the liberation of 1 pmol p-nitrophenoll min from the p-nitrophenyl glycoside. The previously reported [2] assay method using pig gastric mucus glycoprotein (PGM) as the substrate was slightly modified. The liberated monosaccharides and oligosaccharides by endo-GalNAc-ase-S were analyzed by TLC.…”

Section: Methodsmentioning

confidence: 99%

“…The crude enzyme was prepared by 80% (mass/vol.) ammonium sulfate precipitation, gel chromatofocusing and DEAE-Toyopearl as previously described [2]. The enzyme was further purified by N-baminopheny1)-oxamic-acid-agarose chromatography, the previously reported [29] experimental conditions being slightly modified.…”

Section: Methodsmentioning

confidence: 99%

“…(endo-GalNAc-ase-A) [6], which are both commercially available, and can hydrolytically cleave the a-N-acetylgalactosaminyl linkages of only Galpl-3Gal-NAcal-Ser/Thr in glycopeptides and glycoproteins to liberate the disaccharide moiety. The endo-GalNAc-ase-S has been partially purified, and its specific activity, molecular mass and some characteristics have been reported [2], but its action on the sialoglycoprotein has not been examined at all. Fetuin, leukosialin, glycophorin A, submaxillary mucin, intestinal mucin etc.…”

mentioning

confidence: 99%

“…The liberated oligosaccharides from the fetuin were pyridylaminated, and their structures were determined by HPLC and 600-MHz 'H-NMR spectroscopy. A part of the described purification method of endo-GalNAc-ase-S [2] was modified, and synthetic glycosides containing a p-nitrophenyl (Np) group Gal~l-3(GlcNAc~l-6)GalNAc-Np] were used for the screening assay of endo-GalNAc-ase-S activity in the process of the enzyme preparation. In the present study, it was found that endoGalNAc-ase-S could liberate sialyl oligosaccharides from the fetuin, and the susceptibility of the sialyl oligosaccharides in the fetuin to endo-GalNAc-ase-S was compared with the molar ratio of the sialyl oligosaccharides obtained with alkaline sodium borohydride treatment.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy

Ishii-Karakasa

Iwase

Hotta

1997

European Journal of Biochemistry

View full text Add to dashboard Cite

The endo-a-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-I 1242 (endo-GalNAc-ase-S) hydrolyzed the 0-glycosidic linkage between GalNAc and Ser (Thr) in fetuin, liberating oligosaccharides. The 0-linked oligosaccharides liberated from the fetuin with endo-GalNAcase-S were pyridylaminated following fractionation on a Bio-Gel P-4 column. The structure of the pyridylaminated 0-linked oligosaccharides from fetuin has been determined by reverse-phase HPLC and 600-MHz 'H-NMR spectroscopy. The chemical shifts and the coupling constants of pyridylaminated (PA) NeuAca2-3GalpI -3GalNAc were refined by computer simulation of the spectrum. The structures of NeuAca2-3Ga1/31-3(NeuAca2-6)GalNAc-PA and NeuActr2-3GaljZ1-3(NeuAca2-3Gal~l-4GlcNAc~1-6)GalNAc-PA were determined by their structural reporter groups.Keywords: 0-linked sugar chain; 0-glycosidase; NMR; HPLC ; fetuin.The endo-a-N-acetylgalactosaminidase from a culture medium of Streptomyces sp. OH-I 1242 (endo-GalNAc-ase-S) was found to be capable of liberating not only Galpl-3GalNAc but other larger neutral oligosaccharides through the hydrolysis of the 0-glycosidic linkage between the GalNAc and Ser(Thr) residues [I, 21. The substrate specificity of endo-GalNAc-ase-S was different from that of endo-a-N-acetylgalactosaininidase (endoGalNAc-ase), i.e. 0-glycanase, from Diplococcus pneumoniae (endo-GalNAc-ase-D [3 -51) and endo-a-N-acetylgalactosaminidase from Alcaligenes sp. (endo-GalNAc-ase-A) [6], which are both commercially available, and can hydrolytically cleave the a-N-acetylgalactosaminyl linkages of only Galpl-3Gal-NAcal-Ser/Thr in glycopeptides and glycoproteins to liberate the disaccharide moiety. The endo-GalNAc-ase-S has been partially purified, and its specific activity, molecular mass and some characteristics have been reported [2], but its action on the sialoglycoprotein has not been examined at all. Fetuin, leukosialin, glycophorin A, submaxillary mucin, intestinal mucin etc. [7-131 are sialoglycoproteins, which have 0-linked sialyl oligosaccharide5. The structure of the oligosaccharide moiety of these sialoglycoproteins has been partially clarified, and changes in the sialyl oligosaccharide with differentiation, malignant transformation and immunodeficiency have been extensively studied [7,[14][15][16][17][18][19][20][21]. To investigate the oligosaccharide structure of the mucin-type glycoproteins, the O-glycoside linkage was chemically cleaved through weak alkalinity obtained in the p-elimination reactions, and the liberated oligoCorrespondence to K. Hotta, Department of Biochemistry, School of Medicine, Kitasato University, 1-15-1, Kitasato, Sagamiharashi, Kanagawa, Japan 228 F u r +81 427 78 9372. Abbreviutions. PGM, pig gastric mucus glycoprotein; Np, p-nitrophenyl ; PA, pyridylaminated; endo-GalNAc-ase, endo-a-N-acetylgalactosaminidase.Enzymes. Endo-u-N-acetylgalactosaminidase (EC 3.2.1.97) ; sialidase (EC 3.2.1.18).saccharide-alditols were investigated by 'H-NMR spectroscopy [22]. The enzymic cleavage of the 0-glycosidi...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy

Ishii-Karakasa

Iwase

Hotta

1997

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

Mass spectrometry for protein sialoglycosylation

Zhang

Wang

et al. 2017

Mass Spectrometry Reviews

View full text Add to dashboard Cite

Sialic acids are a family of structurally unique and negatively charged nine-carbon sugars, normally found at the terminal positions of glycan chains on glycoproteins and glycolipids. The glycosylation of proteins is a universal post-translational modification in eukaryotic species and regulates essential biological functions, in which the most common sialic acid is N-acetyl-neuraminic acid (2-keto-5-acetamido-3,5-dideoxy-D-glycero-D-galactononulopyranos-1-onic acid) (Neu5NAc). Because of the properties of sialic acids under general mass spectrometry (MS) conditions, such as instability, ionization discrimination, and mixed adducts, the use of MS in the analysis of protein sialoglycosylation is still challenging. The present review is focused on the application of MS related methodologies to the study of both N- and O-linked sialoglycans. We reviewed MS-based strategies for characterizing sialylation by analyzing intact glycoproteins, proteolytic digested glycopeptides, and released glycans. The review concludes with future perspectives in the field.

show abstract

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Reuter¹,

Gabius²

1999

Cellular and Molecular Life Sciences (CMLS)

214

141

View full text Add to dashboard Cite

Protein and lipid glycosylation is a ubiquitous phenomenon. The task of cataloguing the great structural variety of the glycan part has demanded considerable efforts over decades. This patient endeavor was imperative to discern the inherent rules of glycosylation which cannot affirm assumptions on a purely coincidental nature of this type of protein and lipid modification. These results together with theoretical considerations uncover a salient property of oligosaccharides. In comparison with amino acids and nucleotides, monosaccharides excel in their potential to serve as units of hardware for storing biological information. Thus, the view that glycan chains exclusively affect physiochemical properties of the conjugates is indubitably flawed. This original concept has been decisively jolted by the discovery of endogenous receptors (lectins) for distinct glycan epitopes which are as characteristic as a fingerprint or a signature for a certain protein (class) or cell type. Recent evidence documents that these binding proteins are even endowed with the capacity to select distinct low-energy conformers of the often rather flexible oligosaccharides, granting entry to a new level of regulation of ligand affinity by shifting conformer equilibria. The assessment of the details of this recognition by X-ray crystallography, nuclear magnetic resonance spectroscopy, microcalorimetry and custom-made derivatives is supposed to justify a guarded optimism in satisfying the need for innovative drug design in antiadhesion therapy, for example against viral or bacterial infections and unwanted inflammation. This review presents a survey of the structural aspects of glycosylation and of evidence to poignantly endorse the notion that carrier-attached glycan chains can partake in biological information transfer at the level of cell compartments, cells and organs.

show abstract

S1.33 Partial purification and characterization of an endo-?-N-acetylgalactosaminidase from the culture medium ofStreptomyces Sp. OH-11242

Cited by 5 publications

References 0 publications

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy

Mass spectrometry for protein sialoglycosylation

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Contact Info

Product

Resources

About

S1.33 Partial purification and characterization of an endo-?-N-acetylgalactosaminidase from the culture medium ofStreptomyces Sp. OH-11242

Cited by 5 publications

References 0 publications

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz 1H‐NMR Spectroscopy

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz 1H‐NMR Spectroscopy

Mass spectrometry for protein sialoglycosylation

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Contact Info

Product

Resources

About

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy

Structural Determination of the O‐Linked Sialyl Oligosaccharides Liberated from Fetuin with Endo‐α‐N‐Acetylgalactosaminidase‐S by HPLC Analysis and 600‐MHz ¹H‐NMR Spectroscopy