Purification and quantitation of a rat plasma selenoprotein distinct from glutathione peroxidase using monoclonal antibodies.
Studies with 75Se have shown the existence of a rat plasma selenoprotein in addition to glutathione peroxidase. Because the function of the protein is not known, it has been referred to as selenoprotein P. A partially purified preparation was used to produce a monoclonal antibody to selenoprotein P. The antibody did not bind glutathione peroxidase as evidenced by its failure to remove glutathione peroxidase activity from rat plasma by immunoprecipitation. An immunoaffinity column was prepared with the monoclonal antibody, and selenoprotein P was purified 1270-fold from rat plasma in a two-step procedure. The purified selenoprotein P migrated in a single band with an Mr of 57,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Autoradiography demonstrated that this band contained 75Se when the protein was purified from rats which had received 75SeO2-(3). A competitive radioimmunoassay for selenoprotein P was developed. The selenoprotein P concentration in plasma of selenium-replete rats was determined with this assay to be 51 +/- 3.7 micrograms/ml. It was less than 5 micrograms/ml in plasma from selenium-deficient rats. Injection of 50 micrograms of selenium into selenium-deficient rats caused an increase in selenoprotein P from less than 10% of control to 52% of control in 6 h. Plasma glutathione peroxidase activity increased only from 2.2 to 3.1% of control. These experiments demonstrate that rat plasma contains a selenoprotein distinct from glutathione peroxidase. The concentration of this selenoprotein is depressed in selenium deficiency, as is glutathione peroxidase activity, but selenoprotein P increases more rapidly when selenium is supplied than does glutathione peroxidase activity.
Yang
J G
JG
Department of Medicine, University of Texas Health Science Center, San Antonio 78284.
Morrison-Plummer
J
J
Burk
R F
RF
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification; D005979:Glutathione Peroxidase / Q000097:blood*; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D011506:Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D012643:Selenium / Q000097:blood*; D051149:Selenoprotein P; D051140:Selenoproteins
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins; D011506:Proteins; D051149:Selenoprotein P; D051140:Selenoproteins; D005979:Glutathione Peroxidase; D012643:Selenium