Plasma enhances AOC3 activity from Zinc-α2-glycoprotein is an inhibitor of amine oxidase copper-containing 3

2019-07-22T08:12:20Z (GMT) by Matthias Romauch
Supplemental Fig. 1 A, B and C, [14C]-benzylamine assay: Different volumes of murine wt, AOC3 k.o. and ZAG k.o. plasma were tested for amine oxidase activity. In parallel, the same volumes were tested in the presence of the highly selective AOC3 inhibitor LJP1586. D, [14C]-benzylamine assay: recombinant AOC3 (50 ng) activity in the presence of plasma of wt, AOC3 k.o and ZAG k.o mice (male, C57Bl/6 genetic background). Supplemental Fig. 2 A, [14C]-benzylamine assay: Recombinant AOC3 (50 ng) activity in the presence of BSA (bovine serum albumin; fatty acid-free). B, [14C]-benzylamine assay: Comparison of AOC3 activity in wt and ZAG k.o. plasma. For basal plasma AOC3 activity (i.e. without recombinant AOC3, but with 50 ng GST), 50 µl (final concentration 5 mg/ml) of plasma with and without BSA (final concentration 2 mg/ml) were tested. In parallel, the same conditions were tested in the presence of AOC3 (50 ng). C, [14C]-benzylamine assay: AOC3 activity of ZAG-IEX fractions and IEX-fractions lacking ZAG (Fig. 4, B) without addition of recombinant AOC3. AOC3 was replaced with GST (50 ng). D, WB: Plasma membrane proteins from gonadal white adipose tissue (1 µg) of wt and ZAG k.o. mice (four per genotype) were separated by SDS-PAGE and probed using α-AOC3 antibody.