title: Purification of alpha1-antichymotrypsin from human plasma with recombinant M. catarrhalis ubiquitous surface protein A1.
abstract: Alpha 1-antichymotrypsin (ACT) inhibits chymotrypsin-like enzymes, particularly neutrophil cathepsin G. Moreover, ACT in its native form suppresses chemotaxis of neutrophils and decreases neutrophil production of superoxide radicals. We recently showed that Moraxella catarrhalis ubiquitous surface protein (Usp) A1 is able to specifically bind ACT in the context of a novel virulence mechanism. In this study, we report that recombinant UspA1(557-704) coupled to CNBr-Sepharose can be used in a simple one-step purification of ACT from human plasma. UspA1(557-704)-purified ACT remains intact and active as shown by binding to M. catarrhalis and a chymotrypsin inhibition assay. The novel method for ACT isolation from plasma has important advantages in simplicity and time as compared to conventional methods.
authors.affiliation: Medical Microbiology, Department of Laboratory Medicine, Malmö University Hospital, Lund University, SE-205 02 Malmö, Sweden.
authors.forename: Taras
authors.initials: T
authors.lastname: Manolov
authors.forename: Arne
authors.initials: A
authors.lastname: Forsgren
authors.forename: Kristian
authors.initials: K
authors.lastname: Riesbeck
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D001936:Moraxella catarrhalis / Q000737:chemistry*; D000514:alpha 1-Antichymotrypsin / Q000097:blood / Q000302:isolation & purification*
chemical_list: D001425:Bacterial Outer Membrane Proteins; C089675:UspA protein, Moraxella catarrhalis; D000514:alpha 1-Antichymotrypsin; D002918:Chymotrypsin