Substrates of Hageman factor. I. Isolation and characterization of human factor XI (PTA) and inhibition of the activated enzyme by alpha 1-antitrypsin.
Unactivated partial thromboplastin antecedent (PTA) has been purified by sequential chromatography of plasma on quaternary aminoethyl Sephadex, sulphoprophyl Sephadex, Sephadex G-150, and passage over an anti-IgG immunoadsorbant. The preparation gave a single band after alkaline disc gel electrophoresis, sodium dodecyl sulfate (SDS) gel electrophoresis and isoelectric focusing in acrylamide gels and was found to have a mol wt of 175,000 by gel filtration, 163,000 by SDS gel electrophoresis, and an isoelectric point of 8.8-9.4 (peak 9.0-9.1). Pre-PTA was activated directly by activated Hageman factor or by Hageman factor prealbumin fragments. Its coagulant activity was inhibited by DFP, soybean trypsin inhibitor and trasylol but not by lima bean trypsin inhibitor or ovomucoid trypsin inhibitor indicating that activated PTA possesses the same inhibition profile utilizing these reagents as does plasma kallikrein. A major plasma inhibitor of activated PTA was found to be a 65,000 mol wt alpha-globulin which was isolated free of alpha(1)-chymotrypsin inhibitor, inter alpha-trypsin inhibitor, alpha(2)-macroglobulin, and the other known inhibitors of activated PTA, the activated first component of complement (C1 INH), and antithrombin III. Its physicochemical properties were identical to alpha(1)-antitrypsin, and it was absent in alpha(1)-antitrypsin-deficient plasma thereby identifying this PTA inhibitor as alpha(1)-antitrypsin.
D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004586:Electrophoresis; D004589:Electrophoresis, Disc; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism; D005172:Factor XI / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D005174:Factor XII / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D007610:Kallikreins / Q000378:metabolism; D008970:Molecular Weight; D011498:Protein Precursors / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D000515:alpha 1-Antitrypsin / Q000494:pharmacology*
D004792:Enzyme Precursors; D011498:Protein Precursors; D000515:alpha 1-Antitrypsin; D012967:Sodium Dodecyl Sulfate; D005174:Factor XII; D005172:Factor XI; D007610:Kallikreins