Rodent tissue alpha-L-fucosidases: analysis of brain and spleen isoforms and characterization of the purified hamster liver enzyme.
1. Isoelectric focusing of brain and spleen alpha-L-fucosidases from four rodents (rat, mouse, guinea-pig, hamster) indicated that only mouse and hamster tissues contained isoforms with significant amounts of activity above pI 7.0. 2. Hamster liver alpha-L-fucosidase was purified approximately 57,000-fold in 80% yield (to a final specific activity of 24,700 nmol/min/mg protein) by affinity chromatography on agarose-epsilon-aminocaproyl-fucosamine. 3. SDS-PAGE analysis of hamster liver alpha-L-fucosidase indicated the presence of one to two closely-spaced subunits at 56 and 60 kDa. Western blotting analysis indicated the hamster enzyme was recognized by polyclonal antibodies but not by a monoclonal antibody (both antibodies prepared against human liver alpha-L-fucosidase). 4. The pH-activity curve of hamster liver alpha-L-fucosidase is broad with an optimum centered around pH 6.8 and with high activity between pH 5.5 and 7.5.
Shoarinejad
F
F
Department of Chemistry, Lehigh University, Bethlehem, PA 18015.
Alhadeff
J A
JA
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D002846:Chromatography, Affinity; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008099:Liver / Q000201:enzymology*; D008647:Mesocricetus; D051379:Mice; D051381:Rats; D013154:Spleen / Q000201:enzymology*; D005644:alpha-L-Fucosidase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D005644:alpha-L-Fucosidase