Activated alpha 2-macroglobulin is a principal defensin-binding protein.
Defensins are highly abundant and variably cationic peptides that possess antimicrobial, cytotoxic, and chemoattractant properties and equip mammalian phagocytes for participation in host defense and inflammatory processes. We studied the binding of the human defensin HNP-1 by proteins in plasma and serum and identified activated (F-form) alpha 2-macroglobulin (alpha 2M) as a principal binding protein for HNP-1. In contrast, native (S-form) alpha 2M bound little HNP-1. The binding of HNP-1 by F-form alpha 2M was resistant to salt and boiling in 2% sodium dodecyl sulfate but was ablated by dithiothreitol. Pretreatment of methylamine-activated serum or plasma by iodoacetamide substantially decreased the binding of HNP-1 to alpha 2M, suggesting that thiol groups in activated alpha 2M play a role in defensin binding, possibly by covalently trapping defensins via thiol-disulfide exchange. Western blots of conventionally collected sera showed endogenous defensins complexed with the F-form of alpha 2M, indicating that the generation of defensin-alpha 2M complexes was not limited to the in vitro model of methylamine-activated serum or plasma and radiolabeled HNP-1. Previous studies indicated that native alpha 2M can be converted to its F-form by many proteases, including those released by neutrophils and platelets, and that the F-form is recognized and internalized by specific receptors on macrophages and hepatocytes. Our findings suggest that the alpha 2M system may function as a scavenger of defensins and other peptide mediators in inflamed tissues and may constitute an important mechanism for the regulation and containment of inflammation.
Panyutich
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A
Will Rogers Pulmonary Research Laboratory, University of California, Los Angeles 90024-1736.
Ganz
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T
D001770:Blood Bactericidal Activity; D001798:Blood Proteins / Q000378:metabolism*; D023082:Defensins; D006801:Humans; D066298:In Vitro Techniques; D007460:Iodoacetamide / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D023084:alpha-Defensins; D000511:alpha-Macroglobulins / Q000378:metabolism*
D001798:Blood Proteins; D023082:Defensins; D023084:alpha-Defensins; D000511:alpha-Macroglobulins; C406413:human neutrophil peptide 1; D007460:Iodoacetamide