Purification and properties of human urinary β-D-mannosidase

Raffaella Guadalupi, Maguy Bernard, Antonio Orlacchio, Marie José Foglietti, Carla Emiliani

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Two forms of the lysosomal enzyme β-mannosidase were identified and purified from human urine. The purification strategy employed allowed sufficient quantities of both forms to be obtained for subunit analysis and for further characterizations. The two β-mannosidases were identified as β-mannosidase B and A, in order of their elution from an ion-exchange column. In all samples examined, the A form was predominant, and the B/A ratio was consistently 0.14. The two forms displayed the same optimum pH (i.e., 4.3) and both were retained by a Concanavalin-A Sepharose column, but showed different isoelectric points, molecular masses and subunit compositions. Native- and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses of pure β-mannosidases B and A suggest that active protein B (160 kDa) consists of three subunits, one 75 kDa and two 49 kDa subunits. Protein A is smaller and appears to be composed of three subunits of 75 kDa, 49 kDa and 37 kDa. Two forms of β-mannosidase, exhibiting a chromatographic behaviour comparable to the urinary farms, were also detected in human kidney. Nevertheless, in this tissue their relative distribution was different, the B/A ratio being 19.

Original languageEnglish
Pages (from-to)9-16
Number of pages8
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1293
Issue number1
DOIs
Publication statusPublished - Mar 7 1996

Fingerprint

Mannosidases
Purification
Ion Exchange
Isoelectric Point
Staphylococcal Protein A
Molecular mass
Electrophoresis
Sodium Dodecyl Sulfate
Farms
Polyacrylamide Gel Electrophoresis
Ion exchange
Urine
Tissue
Kidney
Enzymes
Chemical analysis

Keywords

  • β-Mannosidase
  • Human
  • Kidney
  • Purification
  • Urine

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology
  • Biophysics

Cite this

Purification and properties of human urinary β-D-mannosidase. / Guadalupi, Raffaella; Bernard, Maguy; Orlacchio, Antonio; Foglietti, Marie José; Emiliani, Carla.

In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, Vol. 1293, No. 1, 07.03.1996, p. 9-16.

Research output: Contribution to journalArticle

Guadalupi, Raffaella ; Bernard, Maguy ; Orlacchio, Antonio ; Foglietti, Marie José ; Emiliani, Carla. / Purification and properties of human urinary β-D-mannosidase. In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular. 1996 ; Vol. 1293, No. 1. pp. 9-16.
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