Immunochemical characterization of feline and human N-acetylgalactosamine 4-sulfatase.
Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI; MPS VI) is a disorder which results from a deficiency in the lysosomal associated enzyme N-acetylgalactosamine 4-sulfatase (4-sulfatase). A feline model of human MPS VI has previously been described and provides a system for the evaluation of enzyme replacement therapy protocols. As a preliminary study to human 4-sulfatase enzyme replacement therapy in feline we have compared the immunochemical properties of human and feline 4-sulfatase. By SDS-PAGE the molecular mass of purified feline and human 4-sulfatase were similar under both reducing and nonreducing conditions. There was, however, a detectable conformation difference between human and feline 4-sulfatase indicating some structural variation. Feline 4-sulfatase reacted weakly with a panel of monoclonal antibodies in an immunobinding assay (interacting with 4-sulfatase in free solution), but the same monoclonal antibodies reacted strongly with feline 4-sulfatase in an immunoquantification assay where the feline 4-sulfatase was bound to a polyclonal antibody (which presumably induces a conformation change in the feline 4-sulfatase to closer approximate the structure of human 4-sulfatase). A monoclonal antibody which selectively reacts with human 4-sulfatase has been used to develop an assay suitable for evaluating human 4-sulfatase enzyme replacement in cat tissues.
Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide, Australia.
D A
DA
Brooks
G J
GJ
Gibson
J J
JJ
Hopwood
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D002415:Cats / Q000378:metabolism*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007120:Immunochemistry; D016014:Linear Models; D008970:Molecular Weight; D020051:N-Acetylgalactosamine-4-Sulfatase; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013429:Sulfatases / Q000032:analysis*; D014020:Tissue Extracts / Q000737:chemistry
D000911:Antibodies, Monoclonal; D014020:Tissue Extracts; D013429:Sulfatases; D020051:N-Acetylgalactosamine-4-Sulfatase