AFFINITY CHROMATOGRAPHY OF RABBIT GASTROCNEMIUS ALPHA-L-FUCOSIDASE
Abstract
Abstract: Ct-L-Fucosidase was purified from rabbit gastrocnemius muscle usingagarose-e-aminocaproyl-fucosamine. The isolated enzyme was found to be pure by anumber of different analytical techniques including gel electrophoresis. The purifiedenzyme was found essentially free of other glycosidases. The main characteristics ofthe purified α-L-Fucosidase were determined; pH 6.0-6. 3, KmO.52, Vmax 25.13 nmol/gm protein/min., Ki 0.46. L-fucosq was the only physiological competitiveinhibitor of the enzyme. Ct-L-fucosidase is heat labile and stable for 2-3 months at —20 C in the presence of lOOmM L-fucose. The purified enzyme showed 50,000 Mol.Wt. on SDS-Electrophoresis.References
Tsay, C.G., Dawson, G. and Sung, S.S.J. Structure of the accumulating oligosaccharide in
fucosidosis. J. Biol. Chem. 1976; 251: 5852-5859.
Opheim, D.J. and Touster, O. The purification and characterisation of rat liver lysosomal a-Lfucosidase. J. Biol. Chem. 1977; 252: 739-743.
Khan, J.A. and Lewis, M.H.R. Alpha-L-fucosidase: Purification and Kinetic properties. Pak. J.
Med. Res. 1985; 24 : 153-161.
Davis, B.J. Disc electrophoresis of human serum proteins. Ann. N.Y. Acad. Sci. 1964;
: 382-392.
Lineweaver, H. and Burk, D. Determination of Kinetic parameters. J.Am. Chem. Soc.
; 56: 658-662.
Robinson, D. and Thorpe, R. Affinity chromatography of human liver cr-L-fucosi- dase.
FEBS Lett; 45: 191-193.
Chien, Su-F. and Dawson, G. Purification and properties of two forms of human CV-Lfucosidase. Biochem. Biophys. Acta. 1980; 614: 476-488.
Jain, R.S., Binder, R.L., Levy-Benshimol, A. Buck, C.A. and Warren, L. Purification of
CT-L-fucosidase from various sources by affinity chromatography. J. Chromatog. 1977
; 139: 283-290.