Singlet oxygen ((1)O2) inactivates plasmatic free and complexed alpha2-macroglobulin.
alpha2-macroglobulin (alpha2M) is a broad-spectrum proteinase inhibitor and one of the major plasma proteins in humans. Activated phagocytes (especially granulocytes) generate large amounts of oxidants of the HOCI- and chloramine-type that release the mild nonradical, excited (light-emitting) oxidant singlet oxygen ((1)O2). These oxidants have been shown to inactivate several specific serine protease inhibitors in human blood [e.g., alpha1-antitrypsin or alpha2-antiplasmin (plasmin inhibitor)]. The studies reported here demonstrate that nonradical oxidants also inactivate plasmatic alpha2M. The effective dose for 50% inactivation (ED50) of plasmatic alpha2M is similar to that for plasmatic alpha2-antiplasmin. Chloramines are about 1,000-fold more effective than hydrogen peroxide (ED50)=0.75 micromol chloramine T/50 microl plasma). Serine protease-serine protease inhibitor complexes are resistant to oxidants. In contrast, here it is shown that alpha2-macroglobulin, even after binding to serine proteases is sensitive to oxidation, the captured protease is released from the protease/alpha2M complex. This is the first time that oxidative inactivation of a complexed (i.e., bound to a target protease) human protease inhibitor has be shown. The (1)O2 inhibitors methionine, cysteine, cystine, or ascorbate-in contrast to the oxy-radical scavengers mannitol, superoxide dismutase, or catalase-antagonize the chloramine/NaOCl-mediated inactivation of both uncomplexed and complexed alpha2M. Thus, the oxidant involved here is of nonradicalic nature and has reaction characteristics of (1)O2. For the inhibitory function, critical oxidizable methionines or the internal thiol-ester might be targets for (1)O2. Consequently, alpha2M can also be considered a carrier for proteases, since the alpha2M-complexed proteases regain full activity in an oxidative environment. In local areas of inflammation or thrombolysis, activated phagocytes could create microenvironments of uncontrolled protease activity by generation of (1)O2.
Stief
T W
TW
Institute of Clinical Chemistry and Molecular Diagnostics, Philipps-University, Marburg, Germany. thstief@post.med.uni-marburg.de
Kropf
J
J
Kretschmer
V
V
Doss
M O
MO
Fareed
J
J
D002700:Chloramines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000378:metabolism; D026082:Singlet Oxygen; D000979:alpha-2-Antiplasmin / Q000187:drug effects; D000511:alpha-Macroglobulins / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000378:metabolism
D002700:Chloramines; D011480:Protease Inhibitors; D000979:alpha-2-Antiplasmin; D000511:alpha-Macroglobulins; D026082:Singlet Oxygen; D006861:Hydrogen Peroxide; C030816:chloramine; D010100:Oxygen