Characterization of an anti-beta-galactosidase antibody recognizing a precursor but not a mature enzyme in solution.
A monospecific antibody was raised against a recombinant human 84-kDa beta-galactosidase precursor protein, which had been produced with a baculovirus gene expression system and affinity-purified. It bound to the 64-kDa mature enzyme protein extracted from human fibroblasts and the antigenic recombinant precursor on immunoblotting, but neither the fibroblast-derived mature enzyme nor the 64-kDa mature protein-like tryptic product of the recombinant precursor protein was immunoprecipitated. We conclude that it specifically recognized the precursor but not the mature protein in solution. Immunoprecipitation with this anti-precursor antibody revealed that the precursor protein mainly accounted for the residual enzyme activity in fibroblasts from an adult GM1-gangliosidosis patient, and the mature protein accounted for the activity in fibroblasts from a juvenile GM1-gangliosidosis patient.
Department of Clinical Genetics, Tokyo Metropolitan Institute of Medical Science, Japan.
N
N
Takiyama
K
K
Itoh
A
A
Oshima
H
H
Sakuraba
Y
Y
Suzuki
D000818:Animals; D000906:Antibodies; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000032:analysis* / Q000276:immunology; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G; D008970:Molecular Weight; D009036:Moths; D010446:Peptide Fragments / Q000032:analysis; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D012867:Skin / Q000201:enzymology; D012996:Solutions; D014162:Transfection; D014357:Trypsin; D001616:beta-Galactosidase / Q000032:analysis* / Q000276:immunology*
D000906:Antibodies; D004792:Enzyme Precursors; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011994:Recombinant Proteins; D012996:Solutions; D001616:beta-Galactosidase; D014357:Trypsin