Redox properties of the lipocalin alpha1-microglobulin: reduction of cytochrome c, hemoglobin, and free iron.
alpha1-Microglobulin (alpha1m) is a 26-kDa plasma and tissue glycoprotein. The protein has a heterogeneous yellow-brown chromophore consisting of small unidentified prosthetic groups localized to a free thiol group (C34) and three lysyl residues (K92, K118, and K130) around the entrance to a hydrophobic pocket. It was recently reported that alpha1m can bind heme and that a C-terminally processed form of alpha1m degrades heme. It is shown here that alpha1m has catalytic reductase and NADH-dehydrogenase-like activities. Cytochrome c, nitroblue tetrazolium (NBT), methemoglobin, and ferricyanide were reduced by alpha1m. Comparison of the reduction rates suggests that methemoglobin is a better substrate than cytochrome c, NBT, and ferricyanide. The reactions with cytochrome c and NBT were mediated by superoxide anions since they were inhibited by superoxide dismutase. The addition of the biological electron donors NADH, NADPH, or ascorbate enhanced the reduction rate of cytochrome c approximately 30-fold. Recombinant alpha1m, which has much less chromophore than plasma and urine alpha1m, was a stronger reductant than the latter alpha1m forms. Site-directed mutagenesis of C34, K92, K118, and K130 and thiol group chemistry showed that the C34 thiol group was involved in the redox reaction but relies upon cooperation with the lysyl residues. The redox properties of alpha1m may provide a physiological protection mechanism against extracellularly exposed heme groups and other oxidants.
Allhorn
Maria
M
Department of Cell and Molecular Biology, Lund University, BMC, B14, 221 84 Lund, Sweden.
Klapyta
Anna
A
Akerström
Bo
B
D000510:Alpha-Globulins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D000975:Antioxidants / Q000502:physiology; D001205:Ascorbic Acid / Q000737:chemistry; D045304:Cytochromes c / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005292:Ferricyanides / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D007501:Iron / Q000737:chemistry*; D008706:Methemoglobin / Q000737:chemistry; D008956:Models, Chemical; D009154:Mutation; D009243:NAD / Q000737:chemistry; D009249:NADP / Q000737:chemistry; D009580:Nitroblue Tetrazolium / Q000737:chemistry; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D011485:Protein Binding; D013053:Spectrophotometry
D000510:Alpha-Globulins; D000975:Antioxidants; D005292:Ferricyanides; D006454:Hemoglobins; C016001:alpha-1-microglobulin; D009243:NAD; D009580:Nitroblue Tetrazolium; D009249:NADP; D045304:Cytochromes c; D008706:Methemoglobin; D007501:Iron; D010088:Oxidoreductases; D001205:Ascorbic Acid; C028033:potassium ferricyanide