TY - JOUR
T1 - Rational Design and Expression of a Heparin-Targeted Human Superoxide Dismutase
AU - Boissinot, Maurice
AU - Kuhn, Leslie A.
AU - Lee, Pandora
AU - Fisher, Cindy L.
AU - Wang, Fisher
AU - Hallewell, Robert A.
AU - Tainer, John A.
PY - 1993
Y1 - 1993
N2 - In order to improve the therapeutic effectiveness of human Cu, Zn superoxide dismutase (HSOD) by targeting it to cell surfaces and increasing its circulatory half-life, we have designed and expressed a heparin-binding derivative of HSOD. This design was based on the idea that structurally independent protein units, HSOD and the heparin-binding A+ helix from protein C inhibitor, could be combined with a carefully chosen linker, GlyProGly, to form a stable, bifunctional protein. The chimeric HSOD-GlyProGly-A+ protein was expressed and secreted to the periplasm of E. coli and had normal SOD activity. HSOD-GlyProGly-A+ had a significantly increased retention time relative to wild-type HSOD on a heparin affinity column, indicating that it was successfully targeted to heparin, and this binding was maintained at physiological ionic strength. When administered to mice, HSOD-GlyProGly-A+ had a half-life of ∼15 minutes, twice that of wild-type HSOD. Our rational design approach should be generally applicable to the creation of bifunctional chimeric molecules.
AB - In order to improve the therapeutic effectiveness of human Cu, Zn superoxide dismutase (HSOD) by targeting it to cell surfaces and increasing its circulatory half-life, we have designed and expressed a heparin-binding derivative of HSOD. This design was based on the idea that structurally independent protein units, HSOD and the heparin-binding A+ helix from protein C inhibitor, could be combined with a carefully chosen linker, GlyProGly, to form a stable, bifunctional protein. The chimeric HSOD-GlyProGly-A+ protein was expressed and secreted to the periplasm of E. coli and had normal SOD activity. HSOD-GlyProGly-A+ had a significantly increased retention time relative to wild-type HSOD on a heparin affinity column, indicating that it was successfully targeted to heparin, and this binding was maintained at physiological ionic strength. When administered to mice, HSOD-GlyProGly-A+ had a half-life of ∼15 minutes, twice that of wild-type HSOD. Our rational design approach should be generally applicable to the creation of bifunctional chimeric molecules.
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U2 - 10.1006/bbrc.1993.1038
DO - 10.1006/bbrc.1993.1038
M3 - Article
C2 - 8422249
AN - SCOPUS:0027313589
SN - 0006-291X
VL - 190
SP - 250
EP - 256
JO - Biochemical and biophysical research communications
JF - Biochemical and biophysical research communications
IS - 1
ER -