Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus

J. Hata, M. Dhar, M. Mitra, M. Harmata, F. Haibach, P. Sun, Daniel Smith

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Exoglycosidases modify carbohydrate epitopes on glycoproteins and glycolipids. The N-acetyl-α-D-galactosaminidase from the domestic chicken, Gallus domesticus, is an important exoglycosidase which degrades the human blood group A epitope. This enzyme has never been demonstrably purified or thoroughly characterized. We have developed a technique to purify this enzyme to homogeneity. The isolated enzyme has a molecular weight of 49.1 kDa by SDS PAGE and 145.0 kDa by gel filtration. The enzyme is highly selective for PNP-N-acetyl-α-D-galactosaminide and is inactive against other low molecular weight substrates. The enzyme hydrolyzes the terminal N-acetyl-α- D-galactosaminide residues from blood group A2 erythrocytes. Protease activity is below detectable limits. The enzyme has a pH optima of 3.7, a pI of 8.15, is relatively unaffected by ionic strength, and is stable at 4°C.

Original languageEnglish (US)
Pages (from-to)77-86
Number of pages10
JournalBiochemistry International
Volume28
Issue number1
StatePublished - 1992
Externally publishedYes

Fingerprint

Hexosaminidases
Purification
Chickens
Enzymes
Glycoside Hydrolases
Blood Group Antigens
Epitopes
Molecular Weight
Molecular weight
Glycolipids
Ionic strength
varespladib methyl
Osmolar Concentration
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Glycoproteins
Peptide Hydrolases
Erythrocytes
Gels
Carbohydrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hata, J., Dhar, M., Mitra, M., Harmata, M., Haibach, F., Sun, P., & Smith, D. (1992). Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus. Biochemistry International, 28(1), 77-86.

Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus. / Hata, J.; Dhar, M.; Mitra, M.; Harmata, M.; Haibach, F.; Sun, P.; Smith, Daniel.

In: Biochemistry International, Vol. 28, No. 1, 1992, p. 77-86.

Research output: Contribution to journalArticle

Hata, J, Dhar, M, Mitra, M, Harmata, M, Haibach, F, Sun, P & Smith, D 1992, 'Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus', Biochemistry International, vol. 28, no. 1, pp. 77-86.
Hata J, Dhar M, Mitra M, Harmata M, Haibach F, Sun P et al. Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus. Biochemistry International. 1992;28(1):77-86.
Hata, J. ; Dhar, M. ; Mitra, M. ; Harmata, M. ; Haibach, F. ; Sun, P. ; Smith, Daniel. / Purification and characterization of N-acetyl-α-D-galactosaminidase from Gallus domesticus. In: Biochemistry International. 1992 ; Vol. 28, No. 1. pp. 77-86.
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