Copper binding to mouse liver S-adenosylhomocysteine hydrolase and the effects of copper on its levels.
The dissociation constant and stoichiometry of copper binding to mouse liver S-adenosylhomocysteine hydrolase (SAHH) was determined as part of characterizing the possible roles of SAHH in copper metabolism. Copper (64Cu(II)) binding was measured by an ultrafiltration method in the presence of EDTA as a competing ligand. The KD was 3.9 +/- 0.7 x 10(-16) M, and the stoichiometry was one g atom of copper per 48-kDa subunit. Western blots indicated that the liver contains approximately 12 times more SAHH than the kidney, which in turn contains approximately 5 times more SAHH than the brain. The high concentration and copper affinity of SAHH in the liver may contribute to the liver's ability to preferentially accumulate copper, and the low levels of SAHH in the brain may contribute to the sensitivity of the brain to copper deficiency. The effects of genetic defects of copper metabolism and copper deficiency on SAHH were also determined. Normal SAHH levels were detected in brindled mouse liver, kidney, and brain. However, SAHH from brindled mouse liver eluted abnormally from phenyl Superose columns implying an effect of the brindled mouse defect on SAHH protein structure. Hepatic cytosols from the toxic milk mouse contained approximately 42% the amount of SAHH detected in controls, and hepatic levels of SAHH were also decreased by approximately 45% in copper-deficient mice. The binding properties of SAHH and the effects of abnormal states of copper metabolism on its levels are consistent with significant roles for SAHH in normal and abnormal copper metabolism. SAHH may have roles in regulating tissue copper levels and the distribution of intracellular copper.
Bethin
K E
KE
Department of Biochemistry, State University of New York at Buffalo 14214, USA.
Cimato
T R
TR
Ettinger
M J
MJ
D043383:Adenosylhomocysteinase; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002850:Chromatography, Gel; D003300:Copper / Q000032:analysis / Q000172:deficiency / Q000378:metabolism* / Q000494:pharmacology*; D003600:Cytosol / Q000201:enzymology; D004790:Enzyme Induction / Q000187:drug effects; D005260:Female; D006867:Hydrolases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008892:Milk; D009928:Organ Specificity; D011485:Protein Binding
D003300:Copper; D006867:Hydrolases; D043383:Adenosylhomocysteinase; C029892:cupric chloride