Oxidized caprine alpha-2-macroglobulin: damaged but not completely dysfunctional.
Caprine alpha-2-macroglobulin (alpha2M) is a broad-spectrum, homotetrameric proteinase inhibitor that can maximally bind a single molecule of proteinase. Inhibition of proteinases by caprine alpha2M results from a series of conformational changes that are initiated by the proteinase and results in physical sequestration of the proteinase within the closed cage-like structure of conformationally altered alpha2M. In a previous study, uric acid-generated superoxide anion was identified as one of the physiologically relevant inactivators of alpha2M S.A. Khan, F.H. Khan [Free. Radic. Res. 34 (2001) 113]. We now demonstrate that hypochlorous acid (HOCl) and, to lesser extent, hydrogen peroxide (H2O2) destroy the antiproteolytic potential of caprine alpha2M. At physiologically attainable concentration, we found that HOCl significantly compromised functional integrity of the inhibitor. High concentrations of H2O2 also partially diminished proteinase inhibitory capacity of alpha2M by a mechanism not involving formation of hydroxyl radicals. For hydrogen peroxide, catalase completely protected alpha2M activity while the ability to protect the inhibitor from HOCl-induced inactivation was limited by availability of albumin. Structure function analysis demonstrated that oxidized caprine inhibitor, unlike its human counterpart, retained its tetrameric configuration as well as its characteristic ability to undergo major conformational change upon trypsinization. It is proposed that inhibition of alpha2M activity may be due to oxidation of essential residues of the inhibitor and/or structural rearrangement of the subunits.
Khan
Shakil A
SA
Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh 202002, India.
Khan
Fahim H
FH
D000818:Animals; D016166:Free Radical Scavengers / Q000378:metabolism; D006041:Goats; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism; D016877:Oxidants / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D000511:alpha-Macroglobulins / Q000737:chemistry / Q000378:metabolism*
D016166:Free Radical Scavengers; D016877:Oxidants; D011480:Protease Inhibitors; D000511:alpha-Macroglobulins; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide