Purification and biochemical characterization of a novel copper, zinc superoxide dismutase from liver of camel (Camelus dromedarius): An antioxidant enzyme with unique properties

Chafik A., Essamadi A., Celik S., MAVİ A.

Bioorganic Chemistry, cilt.86, ss.428-436, 2019 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 86
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.bioorg.2019.02.024
  • Dergi Adı: Bioorganic Chemistry
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.428-436
  • Anahtar Kelimeler: Copper, zinc superoxide dismutase, Camel, Camelus dromedarius, Purification, Biochemical characterization, Unique properties, Stressful desert conditions, CU,ZN-SUPEROXIDE DISMUTASE, GLUTATHIONE TRANSFERASE, PROTEIN, CEREBROCUPREIN, PEROXIDASE, IDENTIFICATION, METABOLISM, CATALASE, LEAVES, CATTLE
  • Atatürk Üniversitesi Adresli: Evet

Özet

A novel copper, zinc superoxide dismutase (CuZnSOD) was purified to homogeneity from the liver of an animal well adapted to the stressful living conditions of the desert, the camel (Camelus dromedarius). The biochemical properties of camel liver CuZnSOD were examined. The purified enzyme had a native molecular weight of 28 kDa, as judged by gel filtration chromatography, and showed a single band at 27 kDa on SDS-PAGE, indicating that it is a monomeric protein. Optimal activity of the purified enzyme occurred at 43 degrees C and pH 6.0, and the activation energy was 1.42 kJ/mol. CuZnSOD activity was strongly inhibited by beta-ME, DTT, H2O2 and SDS and slightly inhibited by EDTA, NaN3 and PMSF. Al-3+,Al- Ca2+, Cd2+, Mg2+ and Zn2+ stimulated CuZnSOD activity, whereas Ba2+, Co2+, Fe2+ and Ni2+ inhibited it. The purified enzyme contained 0.010 mu g of Cu and 0.69 mu g of Zn per mg of protein. K-m, V-max, k(cat) and k(cat)/K-m values for NBT and riboflavin were 16.27 and 0.16 mu M, 20.85 and 21.54 U/mg, 9.65 and 9.97 s(-1), and 0.59 and 62.33 s(-1) mu M-1, respectively. Camel liver CuZnSOD exhibited unique biochemical properties compared to those of other CuZnSODs, including lower molecular weight with a monomeric structure, higher optimum temperature, very low Ea, very low optimum pH, very low contents of Cu and Zn, and higher affinity, turnover number and catalytic efficiency for riboflavin. These unique properties of camel liver CuZnSOD might be related to the ability of this animal to inhabit stressful desert conditions.

A novel copper, zinc superoxide dismutase (CuZnSOD) was purified to homogeneity from the liver of an animal well adapted to the stressful living conditions of the desert, the camel (Camelus dromedarius). The biochemical properties of camel liver CuZnSOD were examined. The purified enzyme had a native molecular weight of 28 kDa, as judged by gel filtration chromatography, and showed a single band at 27 kDa on SDS-PAGE, indicating that it is a monomeric protein. Optimal activity of the purified enzyme occurred at 43 degrees C and pH 6.0, and the activation energy was 1.42 kJ/mol. CuZnSOD activity was strongly inhibited by beta-ME, DTT, H2O2 and SDS and slightly inhibited by EDTA, NaN3 and PMSF. Al-3+,Al- Ca2+, Cd2+, Mg2+ and Zn2+ stimulated CuZnSOD activity, whereas Ba2+, Co2+, Fe2+ and Ni2+ inhibited it. The purified enzyme contained 0.010 mu g of Cu and 0.69 mu g of Zn per mg of protein. K-m, V-max, k(cat) and k(cat)/K-m values for NBT and riboflavin were 16.27 and 0.16 mu M, 20.85 and 21.54 U/mg, 9.65 and 9.97 s(-1), and 0.59 and 62.33 s(-1) mu M-1, respectively. Camel liver CuZnSOD exhibited unique biochemical properties compared to those of other CuZnSODs, including lower molecular weight with a monomeric structure, higher optimum temperature, very low Ea, very low optimum pH, very low contents of Cu and Zn, and higher affinity, turnover number and catalytic efficiency for riboflavin. These unique properties of camel liver CuZnSOD might be related to the ability of this animal to inhabit stressful desert conditions.