Purification and enzymatic properties of lysyl hydroxylase from fetal porcine skin.
Lysyl hydroxylase from fetal porcine skin is shown to bind in a highly specific manner to aminoethyl-Sepharose 4B. When coupled to ammonium sulfate fractionation and DEAE-cellulose chromatography, chromatography of lysyl hydroxylase preparations on aminoethyl-Sepharose 4B has yielded a highly purified (greater than 95%) preparation of lysyl hydroxylase. The enzyme consists of two subunits with molecular weights of 70 000 and 115 000. The overall recovery of activity was 2.5%, yielding approximately to 3.5 mg of purified enzyme from 900 g of fetal porcine skin. The enzyme is more active at 30 degrees C than at 37 degrees C and has a pH optimum near 8.0. Both catalase and bovine serum albumin are required by the enzyme for maximum activity. The sulfhydryl reagents p-(chloromercuri)-benzoate, N-ethylmaleimide, and iodoacetamide are potent inhibitors of the enzyme, whereas dithiothreitol appears to be an activator.
D000818:Animals; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D005333:Fetus; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D011247:Pregnancy; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000302:isolation & purification / Q000378:metabolism*; D012867:Skin / Q000201:enzymology*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013552:Swine
D013439:Sulfhydryl Reagents; D006899:Mixed Function Oxygenases; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase; D004229:Dithiothreitol