Immunochemical studies on Rhodotorula gracilis D-amino acid oxidase.
Polyclonal antibodies were prepared from rabbit sera after immunization with holo- and apo-D-amino acid oxidase purified from R. gracilis. Both anti-holo- and anti-apoenzyme IgG fractions (as well as affinity-purified IgG) were highly specific: in blot-transfer analyses after SDS-PAGE only a 39 kDa band, corresponding to enzyme monomer, was recognized even in the partially purified yeast extract. No cross-reaction was detected with pig kidney D-amino acid oxidase. As a difference from the mammalian enzyme, yeast D-amino acid oxidase anti-holo- and anti-apoenzyme IgGs had different properties in inactivation and precipitation experiments, indicating the existence of different antigenicity sites related to the FAD-binding domain in the enzyme.
Pollegioni
L
L
Department of General Physiology and Biochemistry, University of Milano, Italy.
Simonetta
M P
MP
D015153:Blotting, Western; D003605:D-Amino-Acid Oxidase / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D007150:Immunohistochemistry; D011233:Precipitin Tests; D012248:Rhodotorula / Q000201:enzymology*; D013379:Substrate Specificity
D003605:D-Amino-Acid Oxidase