Human prolyl hydroxylase. Purification, partial characterization and preparation of antiserum to the enzyme.
Prolyl hydroxylase was purified from human foetal skin and from a mixture of human foetal tissues by the affinity chromatography procedure using poly(L-proline). The enzyme from both sources was pure, when examined by polyacrylamide gel electrophoresis, as a native protein or in the presence of sodium dodecylsulphate, and enzyme activity recovery varied from 38% to 70% with seven enzyme preparations. The enzyme synthesized from 61.0 mumol to 82.7 mumol hydroxyproline mg protein-1 h-1 degrees C with a saturating concentration of (Pro-Pro-Gly)5 as substrate. The molecular weight of the enzyme was identical with that of the chick prolyl hydroxylase when studied by gel filtration, and the molecular weights of the subunits of the enzyme were about 61000 and 64000 as determined by sodium dodecylsulphate-polyacrylamide gel electrophoresis. The amino acid composition of the human enzyme was very similar to that of the chick prolyl hydroxylase. Antisera to human and chick prolyl hydroxylases were prepared in rabbits. A single precipitin line was seen between the antiserum to human prolyl hydroxylase and the human enzyme in double immunodiffusion, and no cross-reactivity was detected between the human chick enzymes by this technique. However, a distinct cross-reactivity was observed between the human and chick enzymes in inhibition experiments.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000937:Antigen-Antibody Reactions; D002645:Chickens; D003429:Cross Reactions; D005260:Female; D005333:Fetus; D007106:Immune Sera; D005779:Immunodiffusion; D008970:Molecular Weight; D011247:Pregnancy; D011393:Procollagen-Proline Dioxygenase / Q000276:immunology / Q000302:isolation & purification*; D012867:Skin / Q000201:enzymology
D000596:Amino Acids; D007106:Immune Sera; D011393:Procollagen-Proline Dioxygenase