Human serum α-l-fucosidase

Jack A. Alhadeff, Aaron Janowsky

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Human serum α-l-fucosidase has been purified 241 200-fold with 35% yield by an affinity Chromatographie procedure utilizing agarose-ε{lunate}-aminocaproyl-fucosamine. Isoelectric focusing of the purified enzyme indicated the presence of several forms, with the form at pI 5.0 comprising the majority of the activity. Assay of the purified α-l-fucosidase showed only trace amounts of contaminating glycosidases present, with β-galactosidase being the largest contamnant (0.5% by activity). Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated the presence of two subunits with very similar molecular weights (56500 and 54000). Using the p-nitrophenyl substrate, the purified serum α-l-fucosidase had an apparent Michaelis constant of 0.52 mM and a broad pH optimum centered around pH 4.8 with a second, minor optimum at pH 6.1. Gel filtration on Sepharose 6-B indicated an apparent molecular weight of 296 000 ± 30 000. Preincubation with antibodies made previously against purified liver α-l-fucosidase led to quantitative immuno precipitation of the purified serum α-l-fucosidase. Assay of the purified serum α-l-fucosidase for sialic acid indicated the presence of 1.7 μg sialic acid per 100 μg enzyme, about twice that previously found for the purified liver enzyme.

Original languageEnglish (US)
Pages (from-to)133-140
Number of pages8
JournalClinica Chimica Acta
Volume82
Issue number1-2
DOIs
StatePublished - Jan 2 1978
Externally publishedYes

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alpha-L-Fucosidase
Serum
N-Acetylneuraminic Acid
Liver
Sepharose
Assays
Enzymes
Molecular Weight
Molecular weight
Galactosidases
Glycoside Hydrolases
Isoelectric Focusing
Electrophoresis
Sodium Dodecyl Sulfate
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Gels
Antibodies
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Human serum α-l-fucosidase. / Alhadeff, Jack A.; Janowsky, Aaron.

In: Clinica Chimica Acta, Vol. 82, No. 1-2, 02.01.1978, p. 133-140.

Research output: Contribution to journalArticle

Alhadeff, Jack A. ; Janowsky, Aaron. / Human serum α-l-fucosidase. In: Clinica Chimica Acta. 1978 ; Vol. 82, No. 1-2. pp. 133-140.
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