Williams trait. Human kininogen deficiency with diminished levels of plasminogen proactivator and prekallikrein associated with abnormalities of the Hageman factor-dependent pathways. R W Colman, … , J V Pierce, A P Kaplan J Clin Invest. 1975;56(6):1650-1662. https://doi.org/10.1172/JCI108247. Research Article An asymptomatic woman (Ms. Williams) was found to have a severe abnormality in the surface-activated intrinsic coagulation, fibrinolytic, and kinin-generating pathways. Assays for known coagulation factors were nromal while Fletcher factor (pre-kallikrein) was 45%, insufficient to account for the observed markedly prolonged partial thromboplastin time. Plasminogen proactivator was present at 20% of normal levels and addition of highly purified plasminogen proactivator containing 10% plasminogen activator partially corrected the coagulation and fibrinolytic abnormalities but not the kinin- generating defect. This effect was due to its plasminogen activator content. In addition, Williams trait plasma failed to convert prekallilrein to lakkilrein or release kinin upon incubation with kaolin. Kininogen antigen was undetectable. When normal plasma was fractionated to identify the factor that corrects all the abnormalities in Williams trait plasma, the Williams factor was identified as a form of kininogen by its behavior on ion exchange chromatography, gel filtration, disc gel electrophoresis, and elution from an anti-low molecular weight kininogen immunoadsorbent. High molecular weight kininogen as well as a subfraction of low molecular weight kininogen, possessed this corrective activity while the bulk of low molecular weight kininogen functioned only as a kallikrein substrate. Kininogen therefore is a critical factor required for the functioning of Hageman factor-dependent coagulation and fibrinolysis and for the activation of prekallikrein. Find the latest version: https://jci.me/108247/pdf Williams Trait HUMAN KININOGEN DEFICIENCY WITH DINI4NISHED LEVELS OF PLASMINOGEN PROACTIVATOR AND PREKALLIKREIN ASSOCIATED WITH ABNORMALITIES OF THE HAGEMAN FACTOR-DEPENDENT PATHWAYS ROBERT W. COLMAN, ANDRANIK BAGDASARIAN, RICHARD C. TALAMO, CHERYL F. ScoTr, MARYA SEAVEY, JORGE A. GUIMARAES, JACK V . PIERCE, and ALLEN P. KAPLAN with the technical assistance of LISSA WEINSTEIN From the Coagulation Unit of the Hematology-Oncology Section, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104; The Department of Pediatrics, Johns Hopkins Hospital, Baltimore, Maryland 21205; The Section on Physiological Chemistry, Hypotension-Endocrine Branch, National Heart and Luing Institute; and The Allergic Diseases Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectiouls Diseases, National Institutes of Health, Bethesda, Maryland 20014 A B S T R A C T An asymptomatic woman (Ms. \Vil- gen by- its behavior on ion exchange chromiiatography, liams) was found to have a severe abnormality in the gel filtration, disc gel electrophoresis, and elution from surface-activated intrinsic coagulation, fibrinolvtic, and an anti-low molecular wveight kininogen immunoad- kinin-generating pathways. Assays for known coagula- sorbent. High molecular weight kininogen, as well as tion factors were normal while Fletcher factor (pre- a subfraction of low molecular weight kininogen, pos- kallikrein) was 45%, insufficient to account for the ob- -sessed this corrective activity while the bulk of low served markedly prolonged partial thromboplastin time. molecular Neight kininogen functioned only as a kalli- Plasminogen proactivator was present at 20% of nor- krein substrate. Kininogen therefore is a critical factor mal levels and addition of highly purified plasminogen required for the functioning of Hageman factor-de- proactivator containing 10% plasminogen activator par- pendent coagulation and fibrinolvsis and for the activa- tially corrected the coagulation and fibrinolytic abnor- tion of prekallikrein. malities but not the kinin-generating defect. This effect was due to its plasminogen activator content. In addi- tion, Williams trait plasma failed to convert prekalli- INTRODUCTION krein to kallikrein or release kiniin upon incubation with ILagemiian factor (Factor XII) is knoNvii to initiate the kaolin. Kininogen antigen was undetectable. When nor- I)lasma coagulation (1), kinin-forming (2), and fibrino- mal plasma was fractionated to identify the factor that lytic pathways of human plasma (3, 4) by activating corrects all the abnormalities in Williams trait plasma. the precursor proteins, plasma thromboplastin antecedent the Williams factor was identified as a form of kinino- (Factor XI) (PTA) 1 (1), prekallikrein (Fletcher fac- tor) (5), and plasminogein proactivator (6), respec- This work was presented at the International Conference to their respective activ e enzymes. Although of the Biology of the Kallikrein-Kinin System in Health and tivelv, Disease, Reston, Va., 22 October 1974. and has appeared in abstract form (Fed. Proc. 1975. 34: 859). 1 Abbreziations used in this paper: CM, carboxymethyl; Address reprint requests to Dr. A. Kaplan at the National DFP, diisopropylfluorophosphate; PTA, plasma thrombo- Institutes of Health, Bethesda, MId. plastin antecedent; PTT, partial thromboplastin time; QAE, Rcceived for pliblicationt 27 Mlay, 1975 anid in rezvised forni quaternary aminoethyl; SDS, sodium dodecyl sulfate; SP, 18 Aufgutst 1975. sulfopropyl; TAMe, tosyl L-arginine methyl ester. 1650 The Journal of Clinical Investigation Volume 56 December 1975-1650-1662 patients with deficiency of Factor XII (Hageman trait) pl!asma wvas discarded. Plasma from Ms. Williams was have no symptoms (7), in vitro testing of their plasmas collected and handled identically. reveals profound defects Plasma utilized for the isolation of Hageman factor frag- in surface-activated coagula- ments, prekallikrein, plasminogen proactivator, Factor XI tion, kinin formation, and fibrinolysis. In contrast, Fac- (pre-PTA), plasminogen, and kininogen was collected in tor XI deficiency in man results in a mild bleeding 0.38% sodium citrate. 3.6 mg of hexadimethrine bromide tendency (8) with a defect in the rate of thrombin in 0.1 ml of 0.15 M saline was added for each 10 ml of formation but no abnormalit) in kallikrein or plasmin blood drawn. The tubes were centrifuged at 900 g for 20 min at 4°C and the plasma was separated with plastic formation. Individuals lacking prekallikrein (9) re- pipettes. Plastic columns and test tubes were utilized semble those with Factor XII deficiency and display, throughout all chromatographic procedures to minimize in addition to the expected absence of kinin formation, a contact activation of Hageman factor. Samples were con- diminished rate of surface-activated coagulation and centrated by ultrafiltration tlhrough a UM-10 membrane (Amicon Corp., Lexington, Mass.). Gel filtration on Sepha- fibrinolysis (10, 11). These latter abnormalities are dex G 150 (6), alkaline disc gel electrophoresis (5), and attributed to a feedback requirement in which kallikrein sodium dodecyl sulfate (SDS) gel electrophoresis (17) activates Factor XII to an enzvme that can then act were performed as previously described. Protein was ap- upon each substrate (11-13). proximated by absorbance at 280 nm, with Am'01% assumed This report describes an uinusual to equal 10, or was determined by micro-Kjeldalhl analysis. patient, Ms. Wil- Activated Hageman factor. Fcr experiments utilizing liams, with profound abnormalities of the Hageman the radioimmunoassay for bradykinin, the large activator factor-dependent pathwavs. who has diminislhed levels and prealbumin fragments derived from Hageman factor of prekallikrein and plasminogen proactivator and un- were prepared by sequential fractionation of plasma with detectable kininogen. High molecular weight kininogen ethanol, isoelectric precipitation, ion exchange chromatog- and a subfraction of raphy utilizing DEAE cellulose and CMI Sephadex, and low molecular weight kininogen gel filtration on Sephadex G 150, as described by Bag- are identified as the corrective factors. dasarian et al. (18). Hageman factor prealbumin fragments utilized in fibrino- METHODS lytic assays and in the bioassay of prekallikrein were puri- fied by chromatography of plasma on QAE Sephadex twice, Bradykinin triacetate (Sandoz Ltd., Basel, Switzerland) Sephadex G 100, SP Sephadex, and elution from alkaline was used as the standard for native bradykinin. Hexadimeth- disc gels after electrophoresis, as previously reported by rine bromide (Aldrich Chemical Co., Inc., Milwaukee, Kaplan et al. (19). After dialysis, the fragments were con- WTis.); tosyl L-arginine methyl ester (TAMe), and agarose centrated and routinely used at 25 ,g/ml. When assessed (Sigma Chemical Co., Inc., St. Louis, Mo.); enzodiffusion functionally, 5 ul of Hageman factor fragment generated fibrin plates, polystyrene dishes for immunodiffusion, and 100 ng of bradykinin after incubation with 0.2 ml fresh streptokinase (Hyland Div., Travenol Laboratories, Inc., plasma for 2 min at 37° C. There was no detectable con- Costa Mesa, Calif.); quaternary aminoethyl (QAE) Sepha- tamination with any of the Hageman factor substrates, dex A-25, sulphopropyl (SP) Sephadex, carboxymethyl plasminogen, plasmin, or kininogen. (CM) Sephadex, Sephadex G 150, and G 200 (Pharmacia Hagcan factor suibstrates. Pre-PTA, prekallikrein, and Fine Chemicals, Inc., Piscataway, N. J.); DEAE 52 cellu- plasminogen proactivator were isolated from the effluent lose (Whatman