Human prolyl hydroxylase. Purification, radioimmunoassay and clinical studies in liver diseases.
Prolyl hydroxylase was purified from human placentae, specific antiserum against it was prepared, and a new radioimmunoassay system employing 125I-labelled enzyme preparation was established. The molecular weight of the placental enzyme was shown to be 320,000 by gel filtration. SDS-polyacrylamide gel electrophoresis showed two bands of unequal intensity having molecular weights of 60,000 and 130,000. Their amino acid compositions were identical to each other, suggesting the polypeptide with a molecular weight of 130,000 might be a dimer of the polypeptide with a molecular weight of 60,000. The new radioimmunoassay established had a sensitivity of the order of 10 ng/ml, indicating it was more sensitive than previous radioimmunoassay employing 3H-labelling method. Clinical studies on patients with liver diseases disclosed that the concentrations of serum immunoreactive prolyl hydroxylase were elevated both in cases of hepatocellular damage and in cases of cholestasis. In cases of hepatocellular damage the enzyme behaved like cytoplasmic enzymes such as glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and lactic dehydrogenase, but in cases of cholestasis it resembled biliary enzymes such as alkaline phosphatase and gamma-glutamyl transpeptidase. This result might be associated with the peculiar location of the enzyme within the cell, in the membrane of rough endoplasmic reticulum.
Nagai
Y
Y
Oka
H
H
D000596:Amino Acids / Q000032:analysis; D004796:Clinical Enzyme Tests / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D005779:Immunodiffusion; D008107:Liver Diseases / Q000175:diagnosis*; D008970:Molecular Weight; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011393:Procollagen-Proline Dioxygenase / Q000032:analysis / Q000302:isolation & purification*; D011863:Radioimmunoassay / Q000379:methods; D012016:Reference Values
D000596:Amino Acids; D011393:Procollagen-Proline Dioxygenase