Four monoclonal antibodies inhibit the recognition of arylsulphatase A by the lysosomal enzyme phosphotransferase.
The critical step in the sorting of lysosomal enzymes is their recognition by a phosphotransferase in the Golgi apparatus. The topogenic sequences responsible for the recognition by this enzyme have so far only been defined for the lysosomal protease cathepsin D. We have generated four monoclonal antibodies directed against lysosomal arylsulphatase A (ASA). These antibodies inhibit the recognition of ASA by the phosphotransferase in vitro and thus define a region of topogenic sequences in the ASA polypeptide. The antibodies do not interfere with the enzymic activity nor with pH-dependent dimerization of ASA. The epitopes recognized by the antibodies have been located in the second quarter of the ASA polypeptide using chimeric mouse-human ASA molecules. Three of the monoclonal antibodies bind to identical or closely adjacent epitopes, which are formed by the interaction of amino acid residues 165-184 and 202-240. The fourth antibody recognizes a different epitope within amino acids 256-265.
Sommerlade
H J
HJ
Department of Biochemistry II, Georg August Universität Göttingen, Federal Republic of Germany.
Hille-Rehfeld
A
A
von Figura
K
K
Gieselmann
V
V
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002553:Cerebroside-Sulfatase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006224:Cricetinae; D000939:Epitopes / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D010770:Phosphotransferases; D002553:Cerebroside-Sulfatase