title: Prothrombin, albumin and immunoglobulin A form covalent complexes with alpha1-microglobulin in human plasma.
abstract: Molecules containing the 33-kDa plasma protein alpha1-microglobulin were isolated from human plasma by anti-(alpha1-microglobulin) affinity chromatography. Five major bands could be seen after electrophoretic separation of the alpha1-microglobulin-containing proteins under native conditions. Immunoblotting demonstrated alpha1-microglobulin in all five bands. Two of these have been described previously: free alpha1-microglobulin and alpha1-microglobulin complexed with IgA (IgA x alpha1-microglobulin). The other three bands were identified as prothrombin alpha1-microglobulin, albumin x alpha1-microglobulin and dimeric alpha1-microglobulin. Prothrombin x alpha1-microglobulin were 1:2 and 1:1 complexes which carried approximately 1% of total alpha1-microglobulin, had molecular masses of about 145 kDa and 110 kDa upon SDS/PAGE and dissociated completely to free alpha1-microglobulin and prothrombin (72 kDa) when reducing agents were added, suggesting that the complexes were stabilized by disulfide bonds. The alpha1-microglobulin molecules did not inhibit cleavage of prothrombin by factor Xa and were bound to the peptides which were released upon activation of prothrombin. Albumin x alpha1-microglobulin, corresponding to 7% of total plasma alpha1-microglobulin, was a mixture between 1:1 and 1:2 complexes, with masses upon SDS/PAGE of approximately 100 kDa and 135 kDa, respectively. Both these complexes dissociated only partially to free alpha1-microglobulin and albumin when reducing agents were added. The albumin x alpha1-microglobulin complexes carried a yellow-brown chromophore similar to free alpha1-microglobulin. The complex-binding to alpha1-microglobulin did not block the fatty-acid-binding ability of albumin. The plasma concentrations of albumin x alpha1-microglobulin and prothrombin x alpha1-microglobulin were estimated to 5.2 mg/l and 1.1 mg/l, respectively.
authors.affiliation: The Section for Molecular Signalling, Department of Cell and Molecular Biology, Lund University, Sweden.
authors.forename: T
authors.initials: T
authors.lastname: Berggård
authors.forename: N
authors.initials: N
authors.lastname: Thelin
authors.forename: C
authors.initials: C
authors.lastname: Falkenberg
authors.forename: J J
authors.initials: JJ
authors.lastname: Enghild
authors.forename: B
authors.initials: B
authors.lastname: Akerström
mesh_terms: D000510:Alpha-Globulins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood / Q000737:chemistry*; D046911:Macromolecular Substances; D011485:Protein Binding; D011516:Prothrombin / Q000737:chemistry* / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000510:Alpha-Globulins; D007070:Immunoglobulin A; D046911:Macromolecular Substances; D012709:Serum Albumin; C016001:alpha-1-microglobulin; D011516:Prothrombin