Targeting SOD by gene and protein engineering and inhibition of free radical injury.
Although oxygen toxicity of tissues can be decreased by a variety of antioxidants and some enzymes, such as SOD and catalase, their protective effect on tissue injury in various diseases are fairly small predominantly because of their unfavorable in vivo behavior. To minimize oxidative stress in various diseases, such as ischemic myocardial injury, circulatory disturbance and corneal inflammation, we synthesized three types of SOD derivatives by gene and protein engineering technique. One type of SOD (SM-SOD covalently linked with hydrophobic anions) circulates bound to albumin with a half life of 6 h and accumulates in tissues whose local pH is decreased. The other type of SOD (AC-SOD covalently linked with long chain fatty acids via the epsilon-amino group of lysyl residues) anchors onto membrane/lipid bilayers of various cells. The last type of SOD (HB-SOD synthesized by constructing a fusion gene coding human CuZn-type SOD and a C-terminal heparin-binding domain) binds to heparin-like proteoglycans on vascular endothelial cell surface. Intravenous administration of either SM-SOD or HB-SOD markedly inhibited postischemic reflow arrhythmias in the rat. When the left anterior descending artery was occluded permanently, about 65% of animals died within 30 min predominantly due to irreversible ventricular fibrillation; the motality of animals decreased to 15% by administering SM-SOD either before or after occlusion. Topically administered AC-SOD bound to the corneal epithelial cell surface and polymorphonuclear leukocytes and efficiently dismutated superoxide radicals at their cell surface. Thus, endotoxin-induced keratitis was inhibited markedly by topical instillation of AC-SOD.(ABSTRACT TRUNCATED AT 250 WORDS)
Inoue
M
M
Department of Biochemistry, Kumamoto University Medical School, Japan.
Watanabe
N
N
Utsumi
T
T
Sasaki
J
J
D000595:Amino Acid Sequence; D000818:Animals; D003300:Copper; D003327:Coronary Disease / Q000188:drug therapy / Q000378:metabolism; D015195:Drug Design; D016166:Free Radical Scavengers; D005609:Free Radicals; D005796:Genes; D005813:Genes, Synthetic; D006207:Half-Life; D006801:Humans; D008969:Molecular Sequence Data; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000517:prevention & control; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism / Q000633:toxicity; D015202:Protein Engineering; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism / Q000493:pharmacokinetics / Q000627:therapeutic use; D015032:Zinc
D016166:Free Radical Scavengers; D005609:Free Radicals; D011993:Recombinant Fusion Proteins; D003300:Copper; D013482:Superoxide Dismutase; D015032:Zinc; D010100:Oxygen