Cloning and characterization of human liver cytosolic β-glycosidase

M. De Graaf, I. C. Van Veen, I. H. Van der Meulen-Muileman, W. R. Gerritsen, H. M. Pinedo, H. J. Haisma

Research output: Contribution to journalArticle

Abstract

Cytosolic β-glucosidase (EC 3.2.1.21) from mammalian liver is a member of the family 1 glycoside hydrolases and is known for its ability to hydrolyse a range of β-D-glycosides, including β-D-glucoside and β-D-galactoside. We therefore refer to this enzyme as cytosolic β-glycosidase. We cloned the cDNA encoding the human cytosolic β-glycosidase by performing PCR on cDNA prepared from total human liver RNA. Specific primers were based on human expressed sequence tags found in the expressed sequence tag database. The cloned cDNA contained 1407 nt with an open reading frame encoding 469 amino acid residues. Amino acid sequence analysis indicates that human cytosolic β-glycosidase is most closely related to lactase phlorizin hydrolase and klotho protein. The enzyme was characterized by using cell lysates of COS-7 cells transfected with a eukaryotic expression vector containing the cDNA. The biochemical, kinetic and inhibition properties of the cloned enzyme were found to be identical with those reported for the enzyme purified from human liver.

Original languageEnglish (US)
Pages (from-to)907-910
Number of pages4
JournalBiochemical Journal
Volume356
Issue number3
DOIs
StatePublished - Jun 15 2001
Externally publishedYes

Fingerprint

Cloning
Glycoside Hydrolases
Liver
Organism Cloning
Complementary DNA
Expressed Sequence Tags
Enzymes
Lactase-Phlorizin Hydrolase
Glucosidases
Amino Acids
Galactosides
Glucosides
Glycosides
Aptitude
COS Cells
Protein Sequence Analysis
Open Reading Frames
RNA
Kinetics
Databases

Keywords

  • β-galactosidase
  • β-glucosidase
  • Glycoside
  • Mammalian

ASJC Scopus subject areas

  • Biochemistry

Cite this

De Graaf, M., Van Veen, I. C., Van der Meulen-Muileman, I. H., Gerritsen, W. R., Pinedo, H. M., & Haisma, H. J. (2001). Cloning and characterization of human liver cytosolic β-glycosidase. Biochemical Journal, 356(3), 907-910. https://doi.org/10.1042/0264-6021:3560907

Cloning and characterization of human liver cytosolic β-glycosidase. / De Graaf, M.; Van Veen, I. C.; Van der Meulen-Muileman, I. H.; Gerritsen, W. R.; Pinedo, H. M.; Haisma, H. J.

In: Biochemical Journal, Vol. 356, No. 3, 15.06.2001, p. 907-910.

Research output: Contribution to journalArticle

De Graaf, M, Van Veen, IC, Van der Meulen-Muileman, IH, Gerritsen, WR, Pinedo, HM & Haisma, HJ 2001, 'Cloning and characterization of human liver cytosolic β-glycosidase', Biochemical Journal, vol. 356, no. 3, pp. 907-910. https://doi.org/10.1042/0264-6021:3560907
De Graaf M, Van Veen IC, Van der Meulen-Muileman IH, Gerritsen WR, Pinedo HM, Haisma HJ. Cloning and characterization of human liver cytosolic β-glycosidase. Biochemical Journal. 2001 Jun 15;356(3):907-910. https://doi.org/10.1042/0264-6021:3560907
De Graaf, M. ; Van Veen, I. C. ; Van der Meulen-Muileman, I. H. ; Gerritsen, W. R. ; Pinedo, H. M. ; Haisma, H. J. / Cloning and characterization of human liver cytosolic β-glycosidase. In: Biochemical Journal. 2001 ; Vol. 356, No. 3. pp. 907-910.
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