Characterization of human residual catalase of an acatalasemic patient by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electrophoretic blotting and immunodetection.
Isoelectric points and subunit sizes of catalases in human blood and human cultured skin fibroblasts from acatalasemic and normal subjects were analyzed by isoelectric focusing in agarose gel and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively, followed by electroblotting to polyvinylidene difluoride membranes for immunodetection. The results indicated that the isoelectric point of residual catalase in the C fraction prepared from acatalasemic erythrocytes was identical with that of catalase prepared from normal erythrocytes. The residual catalase in homogenates of acatalasemic cultured skin fibroblasts also reacted with anticatalase rabbit serum and had an isoelectric point identical with that of normal catalase. Subunit sizes of normal and acatalasemic catalases in the C fractions of erythrocytes were also found to be identical on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by electroblotting and immunoenzymatic amplification. The results indicated no substantial difference in molecular size and charge of catalase proteins between normal and acatalasemic erythrocytes and fibroblasts.
Ogata
M
M
Department of Public Health, Okayama University Medical School, Japan.
Suzuki
K
K
Satoh
Y
Y
D020642:Acatalasia; D001797:Blood Protein Electrophoresis; D002374:Catalase / Q000097:blood / Q000378:metabolism; D002478:Cells, Cultured; D004563:Electrochemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007525:Isoelectric Focusing / Q000379:methods*; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate
D012967:Sodium Dodecyl Sulfate; D002374:Catalase