Original ContributionIn Vivo Oxidative Modification of Erythrocyte Membrane Proteins in Copper Deficiency
Introduction
It is well-known that copper is an essential nutrient for man and animals and that its deficiency in diets can have serious pathological consequences.1, 2, 3, 4 However, the molecular mechanisms that produce the pathology associated with copper deficiency are not completely understood. Copper deficiency can reduce the activities of Cu, Zn-superoxide dismutase (SOD), ceruloplasmin, and catalase,5, 6, 7 all of which are enzymatic components of the antioxidant defense system. Thus, increased susceptibility to oxidative damage may be one of the biological consequences that contributes significantly to the pathology of copper deficiency. Evidence that oxidative damage can occur during copper deficiency is contained in reports of increased lipid hydroperoxide content in liver mitochondria and microsomes from copper-deficient rats[8] and increased susceptibility of tissues from copper-deficient rats to lipid peroxidation.9, 10 Increased production of breath ethane is further evidence that lipid peroxidation is increased in copper-deficient rats.[11] In fact, there are enough reports of oxidative damage or increased susceptibility to oxidative damage during copper deficiency to suggest that copper could be characterized as an antioxidant nutrient.[12]
Because they are exposed to high concentrations of oxygen and to superoxide anions that are constantly generated from the autoxidation of oxyhemoglobin,13, 14 erythrocytes may be targets for oxidative damage during copper deficiency. The intracellular concentration of superoxide anion is normally controlled by superoxide dismutase, an enzyme whose erythrocyte activity is reduced by copper deficiency.15, 16 Concentration of superoxide anion could thus increase and oxidatively modify cell components. A report of increased malondialdehyde content in erythrocytes from copper-deficient rats supports the contention that erythrocytes are oxidatively damaged during copper deficiency.[17]
Although most studies have confined their examination of oxidant damage during copper deficiency to lipid components, damage also may occur to protein components of the erythrocyte membrane. It is known that erythrocyte membrane proteins can be oxidatively crosslinked by malondialdehyde,18, 19, 20 phenylhydrazine,[20] and o-phenanthroline/CuSO4.[21] These findings suggest that erythrocyte membrane proteins are susceptible to oxidative modification and may be damaged in vivo if intracellular conditions favor the production or accumulation of oxidants. Although enzymatic defenses against oxidants are compromised by copper deficiency, it has not been demonstrated that oxidative damage to erythrocyte membrane proteins occurs. Thus, the objective of the present study was to determine if the oxidative stress produced by copper deficiency can damage proteins associated with the erythrocyte membrane.
Section snippets
Animals and Diets
Male, weanling Sprague–Dawley rats (SASCO, Inc., Omaha, NE)2 were housed in individual stainless steel cages in quarters maintained at 22°C and 50% humidity with a 12-h light/dark cycle. Upon arrival at our facility, the rats were fed a purified diet
Results
Rats consuming copper-deficient diet containing 0.5 μg Cu/g for five weeks had lower liver copper concentrations, hemoglobin concentrations, hematocrits, and ceruloplasmin oxidase activity than their counterparts consuming copper-adequate diet containing 5.9 μg Cu/g (Table 1). Four rats consuming the copper-deficient diet died before the experiment was completed. Reduced liver copper concentrations, anemia, and reduced ceruloplasmin activities are characteristic of low-copper status33, 34, 35
Discussion
Because of their physiological role as oxygen carriers, erythrocytes are exposed to continuous oxidative stress and require effective defensive mechanisms to prevent or reduce oxidative damage from endogenously generated reactive oxygen intermediates. Cu, Zn-SOD, which regulates the intercellular concentration of , is an important enzymatic component of the erythrocyte oxidant defense system. Loss of CuZnSOD activity could reduce the dismutation of and increase the
Acknowledgements
This work was supported in part by a Sigma Xi Grant-in-Aid of Research to Thomas P. LaBerge. We gratefully acknowledge the technical assistance of Eric L. Risovi, Thomas J. Matzke, and Anne C. Thomas.
References (54)
Copper in human and animal health
- et al.
Biochemical changes in copper deficiency
J. Nutr. Biochem.
(1990)- et al.
Changes in the activity of the Cu-Zn superoxide dismutase enzyme in tissues of the rat with changes in dietary copper
J. Nutr.
(1979) - et al.
The effects of high dietary zinc and copper deficiency on the activity of copper-requiring metalloenzymes in the growing rat
J. Nutr.
(1984) - et al.
Effect of dietary zinc or copper deficiency on the primary free radical defense system in rats
J. Nutr.
(1988) - et al.
Studies on lipid peroxidation in rat liver by copper deficiency
Int. J. Biochem.
(1981) - et al.
The generation of superoxide radical during the autoxidation of hemoglobin
J. Biol. Chem.
(1972) - et al.
Generation of superoxide radicals during the autoxidation of mammalian oxyhemoglobin
Biochim. Biophys. Acta
(1973) - et al.
Copper deficiency and erythrocuprein (2Cu, 2Zn-superoxide dismutase)
Biochim. Biophys. Acta.
(1976)
Enzymes of oxygen metabolism and lipid peroxidation in erythrocytes from copper-deficient rats
Int. J. Biochem.
Crosslinking of red blood cell membrane proteins induced by oxidative stress in β thalassmia
FEBS Lett.
Polymerization of membrane components in aging red blood cells
Biochem. Biophys. Res. Commun.
Cross-linking the major proteins of the isolated erythrocyte membrane
J. Mol. Biol.
Altered cytoskeletal organization and secretory response of thrombin-activated platelets from copper-deficient rats
J. Nutr.
Effect of copper deficiency on erythrocyte membrane proteins of rats
J. Nutr.
Platelet cytochrome c oxidase activity as an indicator of copper status in rats
Nutr. Res.
Human erythrocyte fractionation in “Percoll” density gradients
Clinica Chimica Acta
The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes
Arch. Biochem. Biophys.
Differential susceptibility of plasma proteins to oxidative modification: Examination by Western blot immunoassay
Free Rad. Biol. Med.
Copper activation of superoxide dismutase in rat erythrocytes
Arch. Biochem. Biophys.
Measurement of protein using bicinchoninic acid
Anal. Biochem.
Anemia, iron storage, and ceruloplasmin in copper nutrition in the growing rat
J. Nutr.
Integral membrane protein interaction with triton cytoskeletons of erythrocytes
Biochim. Biophys. Acta
Human erythrocyte separation according to age on a discontinuous “Percoll” density gradient
Clin. Chim. Acta
The relationship between red cell aging and enzyme activities in experimental animals
Comp. Biochem. Physiol.
Enzymatic changes in rat erythrocytes with increasing cell and donor age: Loss of superoxide dismutase activity associated with increases in catalytically defective forms
Biochem. Biophys. Res. Commun.
Cited by (65)
Higher baseline copper levels are associated with worse outcome in burn patients with overweight and obesity
2022, BurnsCitation Excerpt :Moreover, Cu is a component in antioxidant enzymes such as superoxide dismutase (Cu Zn-SOD), an antioxidant enzyme involved in protecting the body from oxidative stress [19,20]. Cu deficiency is associated with low activity of Cu Zn-SOD [21,22]. Hence, low cellular levels of Cu may increase cellular susceptibility to oxidative damage as well.
Dietary copper affects antioxidant status of shrimp (Penaeus monodon) reared in low salinity water
2022, Aquaculture ReportsCitation Excerpt :Cu is an essential cofactor of the important antioxidant enzyme Cu, Zn-SOD (O'Dell, 1976). Restricting dietary Cu could depress the activity of Cu, Zn-SOD and increase oxidative injury in various terrestrial animals, such as chicken, lamb and goat (Sukalski et al., 1997; Bozkaya et al., 2001; Cheng et al., 2011; Zhang et al., 2012). Similar results were observed in some aquatic species.
Detrimental effects of long-term exposure to heavy metals on histology, size and trace elements of testes and sperm parameters in Kermani Sheep
2021, Ecotoxicology and Environmental SafetyNutrition
2019, The Laboratory RatDeveloping transcriptional profiles in Orbicella franksi exposed to copper: Characterizing responses associated with a spectrum of laboratory-controlled environmental conditions
2017, Aquatic ToxicologyCitation Excerpt :The hsp90 expression profiles generated by Venn et al. (2009) and Schwarz et al. (2013) coupled with this investigation suggest hsp90 is effective over a wide range of concentrations and time-frames. Six transcripts (actin-Symbio, β-actin, tuba3, β-tubulin, β-tub-Z, Spec-α2) are associated with cytomorphology and membrane organization (Sukalski et al., 1997; Fukuda et al., 2002; Takishita et al., 2005; Shearer et al., 2012; Rosic et al., 2015; Garcia et al., 2016). At higher concentrations, both Symbiodinium spp. transcripts exhibit similar patterns of responsiveness (see Fig. 2D), whereas coral genes ae more variable in expression (see Fig. 2B–D).
Potential in vitro antioxidant and protective effects of Mesua ferrea Linn. bark extracts on induced oxidative damage
2013, Industrial Crops and ProductsCitation Excerpt :The antioxidants play a crucial role in scavenging the active free radicals before they attack biologically vital molecules by donating hydrogen atom to maintain the cellular homeostasis. Generally, erythrocytes have been used as a cellular model to investigate oxidative damage, because they are considered as prime targets for free radical attack owing to the presence of both high membrane concentration of polyunsaturated fatty acids (PUFA) and the redox active protein hemoglobin (Hb), which is the potent promoter of ROS (Sadrazadeh et al., 1984; Sukalski et al., 1997). These erythrocytes undergo hemolysis and change in their shape when it get exposed to harsh conditions of H2O2, which indicates the oxidative damage on the erythrocytes.
- 1
US Department of Agriculture, Agricultural Research Service is an equal opportunity/affirmative action employer and all agency services are available without discrimination.