Purification and properties of human brain α L fucosidase

J. A. Alhadeff, Aaron Janowsky

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Human brain α-L-fucosidase has been extracted and the soluble portion has been purified 9388-fold with 25% yield by a two-step affinity chromatographic procedure utilizing agarose-epsilon-aminocaproyl-fucosamine. Isoelectric focusing revealed that all seven isoelectric forms of the enzyme were purified. Trace amounts of eight glycosidases, with hexosaminidase being the largest contaminant (1% by activity) were found in the purified α-L-fucosidase preparation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated the presence of a single subunit of molecular weight 51,000 ± 2500. The purified enzyme has a pH optimum of 4.7 with a suggested second optimum of 6.6. The apparent Michaelis constant and maximal velocity of the purified enzyme with respect to the p-nitrophenyl substrate are 0.44 mM and 10.7 μmol/min/mg protein, respectively. Ag2+ and Hg2+ completely inactivated the enzyme at concentrations of 0.1-0.3 mM. Antibodies made previously against purified human liver α-L-fucosidase cross-reacted with the purified brain α-L-fucosidase and gave a single precipitin line coincident with that from purified liver α-L-fucosidase. From all our studies it appears that at least the soluble portion of brain α-L-fucosidase is identical to human liver α-L-fucosidase.

Original languageEnglish (US)
Pages (from-to)423-427
Number of pages5
JournalJournal of Neurochemistry
Volume28
Issue number2
StatePublished - 1977
Externally publishedYes

Fingerprint

alpha-L-Fucosidase
Purification
Brain
Liver
Enzymes
Hexosaminidases
Precipitins
Glycoside Hydrolases
Isoelectric Focusing
Electrophoresis
Sodium Dodecyl Sulfate
Sepharose
Polyacrylamide Gel Electrophoresis
Molecular Weight
Molecular weight
Impurities
Antibodies
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Purification and properties of human brain α L fucosidase. / Alhadeff, J. A.; Janowsky, Aaron.

In: Journal of Neurochemistry, Vol. 28, No. 2, 1977, p. 423-427.

Research output: Contribution to journalArticle

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