title: Enhancement of potency and stability of human extracellular superoxide dismutase.
abstract: Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant enzymes are considered biomedicine candidates. Among them, extracellular superoxide dismutase (SOD3) had showed prominent efficacy against asthma and inflammation. Despite its advantages as a biomedicine, the difficulty in obtaining large quantity of active recombinant human SOD3 (rhSOD3) has limited its clinical applications. We found that a significant fraction of overexpressed rhSOD3 was composed of the inactive apo-enzyme and its potency against inflammation depended on the rate of metal incorporation. Also, purified rhSOD3 was unstable and lost its activity very quickly. Here, we suggest an ideal preparative method to express, purify, and store highly active rhSOD3. The enzymatic activity of rhSOD3 was maximized by incorporating metal ions into rhSOD3 after purification. Also, albumin or polyethylene glycol prevented rapid inactivation or degradation of rhSOD3 during preparative procedures and long-term storage.
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040; New Drug Development Center, Daegu-Gyungpook Medical Innovation Foundation, Daegu 701-310, Korea.
authors.forename: Sunghwan
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Hae-Young
authors.initials: HY
authors.lastname: Kim
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Jung-Ho
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Jung-Hye
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Won-Kook
authors.initials: WK
authors.lastname: Ham
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Yoon-Jae
authors.initials: YJ
authors.lastname: Jeon
authors.affiliation: Division of Life Science, College of Life Science and Bioengineering, Incheon National University, Incheon 406-772, Korea.
authors.forename: Hara
authors.initials: H
authors.lastname: Kang
authors.affiliation: Department of Dermatology, College of Medicine, The Catholic University of Korea, Seoul 137-040, Korea.
authors.forename: Tae-Yoon
authors.initials: TY
authors.lastname: Kim
mesh_terms: D003300:Copper / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D011092:Polyethylene Glycols / Q000737:chemistry; D055550:Protein Stability; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015032:Zinc / Q000737:chemistry
chemical_list: D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D011092:Polyethylene Glycols; D003300:Copper; C441642:SOD3 protein, human; D013482:Superoxide Dismutase; D015032:Zinc