Purification, properties and influence of dietary copper on accumulation and functional activity of lysyl oxidase in rat skin.
Lysyl oxidase (protein-lysine 6-oxidase; EC 1.4.3.13) is a copper-containing enzyme that functions extracellularly and catalyses the oxidative deamination of peptidyl lysine. Lysyl oxidase was purified 150-175-fold from urea extracts of rat skin and uteri. Features of the enzyme were similar to those reported previously for lysyl oxidase obtained from rat aorta and bovine ligamenture. However, both approximately 40 and approximately 32 kDa polypeptide chains could be isolated from rat skin with apparent lysyl oxidase activity. Antibodies raised in chickens against the approximately 40 kDa form of lysyl oxidase detected the approximately 32 kDa form in immunoblots. Consequently it is inferred that the approximately 32 kDa form of lysyl oxidase is processed from the approximately 40 kDa form of the enzyme. The antibodies were also used to prepare anti(rat lysyl oxidase) affinity columns to facilitate the separation of lysyl oxidase from other proteins in studies to assess the extent to which lysyl oxidase serves as a reservoir for skin copper. At 16 h after an oral dose of copper, as 67Cu, about 6-8% of the total 67Cu incorporated into rat skin was found in association with lysyl oxidase. The lysyl oxidase concentration in rat skin was 2.5-7.5 nmol/g (determined by e.l.i.s.a.). Changing the copper status of rats by feeding a diet deficient in copper did not appear to influence lysyl oxidase accumulation in skin nor the percentage of incorporation of 67Cu in skin as lysyl oxidase. However, when rats were deprived of copper, the functional activity of lysyl oxidase in skin was one-third to one-half the normal values.
Department of Nutrition, College of Agricultural and Environmental Sciences, University of California, Davis 95616-8669.
N
N
Romero-Chapman
J
J
Lee
D
D
Tinker
J Y
JY
Uriu-Hare
C L
CL
Keen
R R
RR
Rucker
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D003300:Copper / Q000172:deficiency / Q000378:metabolism / Q000494:pharmacology*; D003301:Copper Radioisotopes; D004032:Diet; D005260:Female; D008970:Molecular Weight; D008249:Protein-Lysine 6-Oxidase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012867:Skin / Q000201:enzymology*; D014508:Urea; D014599:Uterus / Q000201:enzymology
D003301:Copper Radioisotopes; D003300:Copper; D014508:Urea; D008249:Protein-Lysine 6-Oxidase