| Popis: |
Human A1M (alpha-1-microglobulin) is a reductase and radical- and heme-binding protein with physiological antioxidant protective functions. Recombinant human A1M has been shown to have therapeutic properties in vitro and in vivo, but lacks glycosylation, and shows lower solubility and stability than A1M purified from human plasma. The aims of this work were to 1) use site-directed mutagenesis to improve the physicochemical properties of A1M, 2) demonstrate that the physicochemically improved A1M displays full in vitro cell protective effects as recombinant wild-type A1M (A1M-wt), and 3) show its therapeutic potential in vivo against acute kidney injury (AKI), a disease associated with oxidative stress, using a mouse rhabdomyolysis glycerol-injection model. A novel recombinant A1M-variant (A1M-035) with three amino acid substitutions was constructed, successfully expressed and purified. A1M-035 had improved solubility and stability compared to A1M-wt, and had intact in vitro heme-binding, reductase, antioxidation, and cell protective activities. Both A1M-035 and A1M-wt showed, for the first time, potential in vivo protective effects on acute kidney injury (AKI). It is concluded that the new A1M-035 is a better drug candidate than A1M-wt for treatment of AKI in human patients. |
Full Text from ScienceDirect