title: Human placental lysyl oxidase. Purification, partial characterization, and preparation of two specific antisera to the enzyme.
abstract: Lysyl oxidase from human placentas gave four catalytically active forms on DEAE-cellulose chromatography in 6 M urea. The first tow of these were combined to form pool I and the remaining two to form pool II. Pool I was purified to homogeneity, while the final pool II enzyme usually had one minor contaminant. The molecular weight of both enzyme pools was identical, being about 30,000 by gel filtration in 6 M urea and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. No distinct differences were found between the two pools in amino acid composition, specific activity, or the use of various substrates. Two antisera were prepared, one to the total enzyme protein (pools I and II) and the other to pool I. Both antisera inhibited and precipitated crude placental lysyl oxidase, the two enzyme pools, and crude human skin fibroblast enzyme, there being no differences between the various enzyme forms. Both antisera also stained the two enzyme pools in immunoblotting of denatured proteins. The data suggest that there are no major catalytic, molecular, or immunological differences between the multiple forms of human lysyl oxidase. An antiserum prepared to any of the enzyme forms can, therefore, probably be used to study the total enzyme protein.
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002642:Chick Embryo; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003094:Collagen / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007106:Immune Sera; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D008249:Protein-Lysine 6-Oxidase / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000596:Amino Acids; D007106:Immune Sera; D003094:Collagen; D000594:Amino Acid Oxidoreductases; D008249:Protein-Lysine 6-Oxidase