Biochemical and immunological characterization of a glycosylated alpha-fucosidase from the invertebrate Unio: interaction of the enzyme with its in vivo binding partners

Protein Expression and Purification
S K NadimpalliSuresh Koduru

Abstract

Mammalian alpha-fucosidase (EC 3.2.1.51) is a lysosomal enzyme that catalyzes the removal of fucose residues from glycosphingolipids and its absence in humans results in a rare metabolic disorder called fucosidosis. Among the invertebrates in the molluscs (Unio) two forms of the enzyme have been reported, a 68 kDa non-glycosylated form and a 56 kDa glycosylated form. The glycosylated form has been purified from the seminal fluid of Unio [Biochem. Biophys. Res. Commun. 234 (1997) 54]. In the present study, the 56 kDa glycosylated form has been purified to homogeneity from the whole body tissue of Unio using a series of chromatographic steps. The purified enzyme migrated as a single protein species in 10% SDS-PAGE. Antibodies to the purified enzyme were raised in a rabbit in order to study its biochemical and immunological properties. The purified enzyme is a glycoprotein that exhibits strong binding to Con A-Sepharose gel and can be deglycosylated by PNGase F enzyme suggesting it to be N-glycosylated. The enzyme has been shown to specifically interact with the mannose 6-phosphate receptor protein (MPR 300) purified from goat and Unio. This specific interaction is discussed in view of its possible in vivo binding partners.

References

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Apr 30, 2004·Glycoconjugate Journal·Koduru SureshSiva Kumar Nadimpalli

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Citations

Jul 16, 2008·Glycobiology·Nancy M DahmsJung-Ja P Kim
Mar 9, 2013·Bioscience, Biotechnology, and Biochemistry·Ashapogu Venugopal, Nadimpalli Sivakumar
Dec 14, 2011·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Visa Merino, Nadimpalli Siva Kumar
Dec 25, 2013·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Ashapogu Venugopal, Nadimpalli Siva Kumar
Feb 23, 2020·International Journal of Biological Macromolecules·Rohit Sai Reddy KonadaSiva Kumar Nadimpalli
Jun 15, 2017·International Journal of Biological Macromolecules·A VenugopalMusti J Swamy

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