Modulation of lysyl oxidase by dietary copper in rats.
Lysyl oxidase levels were estimated in rat tissues using an enzyme-linked immunosorption assay (ELISA) and a functional assay standardized against known amounts of purified lysyl oxidase. High concentrations of lysyl oxidase (> or = 150 micrograms/g of tissue or packed cells) were detected in connective tissues, such as tendon and skin. Values for aorta, kidney, lung and liver ranged from 30 to 150 micrograms/g of tissue; values for skeletal muscle and diaphragm were < 30 micrograms/g tissue. Purified rat skin lysyl oxidase catalyzed the release of 50-100 Bq of tritium per micrograms enzyme in assays that used 3H-elastin-rich substrates. In dense connective tissues, good agreement was obtained for the values from ELISA and those derived from measurements of functional activity in aorta, lung, skin and tendon (r2 > 0.9). When egg white-based experimental diets containing 2 or 10 micrograms/g added copper were fed to weanling rats, values for skin lysyl oxidase functional activity in the group fed 2 micrograms/g added copper were one-third to one-half the values for skin lysyl oxidase functional activity in rats fed 10 micrograms/g copper. This reduction in lysyl oxidase activity, however, had minimal effect on indices of collagen maturation in rat skin, e.g., collagen solubility in neutral salt and dilute acid or the levels of acid stable cross-links. Moreover, copper deficiency did not influence the steady-state levels of lysyl oxidase specific mRNA in rat skin or the apparent amounts of lysyl oxidase in rat skin as determined by ELISA. These observations underscore that the concentration of lysyl oxidase is relatively high in dense corrective tissues, and although decreasing dietary copper influences functional activity, there is little apparent effect on the production of lysyl oxidase protein.
Rucker
R B
RB
Department of Nutrition (College of Agriculture and Environmental Sciences), University of California, Davis 95616, USA.
Romero-Chapman
N
N
Wong
T
T
Lee
J
J
Steinberg
F M
FM
McGee
C
C
Clegg
M S
MS
Reiser
K
K
Kosonen
T
T
Uriu-Hare
J Y
JY
Murphy
J
J
Keen
C L
CL
D000284:Administration, Oral; D000818:Animals; D001011:Aorta / Q000201:enzymology / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D003238:Connective Tissue / Q000201:enzymology / Q000378:metabolism; D003300:Copper / Q000008:administration & dosage / Q000172:deficiency / Q000494:pharmacology*; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D008168:Lung / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D008249:Protein-Lysine 6-Oxidase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012867:Skin / Q000201:enzymology / Q000378:metabolism; D013710:Tendons / Q000201:enzymology / Q000378:metabolism
D017931:DNA Primers; D012333:RNA, Messenger; D003300:Copper; D004247:DNA; D008249:Protein-Lysine 6-Oxidase