Antihuman plasma glutathione peroxidase antibodies: immunologic investigations to determine plasma glutathione peroxidase protein and selenium content in plasma.
Plasma glutathione peroxidase (GSHPx) (glutathione: H2O2 oxidoreductase) is a unique selenoglycoprotein. Treatment of this enzyme with glycopeptidase F partially deglycosylates it and establishes the presence of N-linked sugar moieties. Antibodies raised in a rabbit against the purified enzyme from plasma were found to be specific, noninhibitory, and capable of precipitating the enzymatic activity. The antibodies precipitated greater than 90% of the GSHPx activity of normal plasma, thus indicating that the selenoenzyme is the main if not the sole GSHPx activity of plasma. The antibodies did not precipitate RBC GSHPx. A slight cross-reactivity of the antibodies was found with rat plasma GSHPx. A GSHPx activity precipitation assay of normal plasma in the presence of selenium (Se)-deficient plasma indicates that no cross-reactive protein in the Se-deficient plasma interferes with the precipitation of the GSHPx activity from normal plasma. Thus, GSHPx protein as well as activity is deficient in plasma in the absence of Se. Antibodies against GSHPx either from RBCs or from plasma were used to specifically immunoprecipitate most of the GSHPx activity from RBCs or plasma, respectively, in healthy individuals to determine the amount of Se associated with the protein. GSHPx accounts for approximately 15% of the Se in RBCs and 12% of the Se in plasma. Thus, in normal individuals, these proteins account for only a fraction of plasma and RBC Se.
Avissar
N
N
Department of Pediatrics, University of Rochester, School of Medicine and Dentistry, NY 14642.
Whitin
J C
JC
Allen
P Z
PZ
Palmer
I S
IS
Cohen
H J
HJ
D000818:Animals; D000906:Antibodies / Q000032:analysis; D004912:Erythrocytes / Q000032:analysis; D005979:Glutathione Peroxidase / Q000097:blood* / Q000276:immunology; D006026:Glycoside Hydrolases; D006801:Humans; D006868:Hydrolysis; D005779:Immunodiffusion; D007074:Immunoglobulin G / Q000032:analysis; D011233:Precipitin Tests; D011817:Rabbits; D012643:Selenium / Q000097:blood* / Q000172:deficiency
D000906:Antibodies; D007074:Immunoglobulin G; D005979:Glutathione Peroxidase; D006026:Glycoside Hydrolases; D012643:Selenium