Purification of High Molecular Weight Kininogen and the Role of This Agent in Blood Coagulation

Saito, Hidehiko

doi:10.1172/jci108810

Cited by 56 publications

(35 citation statements)

References 29 publications

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“…A crude immunoglobulin fraction was separated from normal rabbit serum or from monospecific rabbit antisera to human HF (17), PTA (18), kallikrein (19), HMWK (13), and plasminogen (Behring, Sommerville, NJ) (20) 10 min, relative to fresh solutions of ellagic acid, using the partial thromboplastin time (PTT) of normal plasma as an indicator. The Sephadex-ellagic acid was washed 6-15 times with 100 ml of buffer, with the supernatant discarded each time after the gel had settled; the last buffer contained 0.02% sodium azide.…”

Section: Methodsmentioning

confidence: 99%

“…This pathway is impaired in plasmas deficient in either PK (6) or HMWK (7). In purified systems, however, while activation of PTA requires the presence of HMWK (8)(9)(10)(11), PK is not a necessary reactant (12,13). One explanation for the difference between plasma and purified systems is that HF is altered during purification, thus obscuring a need for PK. We have reexamined the early steps in the intrinsic pathway to determine whether activation of HF in plasma requires PK or HMWK.…”

mentioning

confidence: 99%

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Interactions among Hageman factor, plasma prekallikrein, high molecular weight kininogen, and plasma thromboplastin antecedent.

Ratnoff¹,

Saito²

1979

Proc. Natl. Acad. Sci. U.S.A.

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To investigate the earliest steps of the intrinsic clotting pathway, Hageman factor (Factor XII) was exposed to Sephadex gels to which ellagic acid had been adsorbed; Hageman factor was then separated from the gels and studied in the fluid phase. Sephadex-ellagic acid-exposed Hageman factor, whether purified or in plasma, activated plasma thromboplastin antecedent, but only when high molecular weight kininogen was present. In the absence of plasma prekallikrein, maximal activation of plasma thromboplastin antecedent was slightly delayed in plasma, a delay not observed with similarly treated purified Hageman factor. Thus, high molecular weight kininogen was needed for expression of Hageman factor's clotpromoting properties and plasma prekallikrein played a minor role in the interaction of ellagic acid-treated Hageman factor and plasma thromboplastin antecedent. Plasma exposed to negatively charged agents clots through reactions of the intrinsic pathway of thrombin formation. The first step in this pathway is activation of Hageman factor (HF, Factor XII). Indirect evidence implies that negatively charged substances bring about a conformational change in HF needed for its clot-promoting activity (1-3). Recent studies suggest that activation of human (4) and bovine (5) HF also involves its proteolytic scission within an internal disulfide loop. Once activated, HF activates plasma thromboplastin antecedent (PTA, Factor XI). The roles of plasma prekallikrein (PK, Fletcher factor) and high molecular weight kininogen (HMWK, Fitzgerald, Williams, Flaujeac, or Reid factor) in the intrinsic pathway are not fully elucidated. This pathway is impaired in plasmas deficient in either PK (6) or HMWK (7). In purified systems, however, while activation of PTA requires the presence of HMWK (8-11), PK is not a necessary reactant (12, 13). One explanation for the difference between plasma and purified systems is that HF is altered during purification, thus obscuring a need for PK. We have reexamined the early steps in the intrinsic pathway to determine whether activation of HF in plasma requires PK or HMWK. Plasma containing HF, or purified HF, acquired clot-promoting properties upon exposure to ellagic acid adsorbed to Sephadex gels. Expression of these properties required the presence of HMWK but not PK. MATERIALS AND METHODSNormal human plasma and plasma from individuals with Hageman trait, Fitzgerald trait (HMWK deficiency), and PTA deficiency were separated from venous blood containing 1/50th vol of 0.5 M sodium citrate buffer (pH 5.0) (14). Fletcher trait (PK deficiency) and Christmas disease (Factor IX deficiency) plasmas were prepared from blood containing 1/9th vol of 0.13 M sodium citrate, and bovine PTA-deficient plasma was from blood containing 1/9th vol of 0.13 M sodium citrate buffer (pH 5.0). A standard pool of normal adult plasmas (14) was said to contain 1 unit/ml each of HF, PK, HMWK, and PTA.Plasmas simultaneously deficient in PTA and other factors were prepared by incubating Fletcher or Fitzgerald trait...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Interactions among Hageman factor, plasma prekallikrein, high molecular weight kininogen, and plasma thromboplastin antecedent.

Ratnoff¹,

Saito²

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…The requirement for cleavage of HMW-kininogen by Factor XII-dependent proteases, before surface adsorption, allows for the temporally ordered proximity of HMW-kininogen, with its complexed prekallikrein and Factor XI, to Factor XIIa. Viewed in this way, diffusion of kallikrein into solution (38) would allow propagation of proteolysis, not only by activation of additional amounts of Factor XII (6), but also by cleavage of HMW-kininogen, in order to maximize its cofactor activity.…”

Section: Effect Of Kallikreinmentioning

confidence: 99%

“…This assessment was derived from the further observation of the lack of prekallikrein adsorption and the diminished Factor XI adsorption in both Factor XII-deficient and HMW-kininogen-deficient plasmas, since these two zymogens (prekallikrein and Factor XI) are transported to a negatively charged surface in complex with HMW-kininogen. The percentage of HMW-kininogen coagulant activity that adsorbed to kaolin closely correlated (r = 0.98, slope Introduction High molecular weight kininogen (HMW)'-kininogen functions as a cofactor in the initiation ofsurface-activated intrinsic blood coagulation by circulating in complexes with either Factor XI (1) or prekallikrein (2), thereby facilitating their optimal orientation on a negatively charged surface (3)(4)(5)(6)(7). Here, the zymogens are converted to their corresponding active enzymes (Factor XIa and kallikrein) by activated Factor XII (8)(9)(10).…”

mentioning

confidence: 99%

Cleavage of human high molecular weight kininogen markedly enhances its coagulant activity. Evidence that this molecule exists as a procofactor.

Scott¹,

Silver²,

Schapira³

et al. 1984

J. Clin. Invest.

130

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Abstract. High molecular weight kininogen (HMW)-kininogen, the cofactor of contact-activated blood coagulation, accelerates the activation of Factor XII, prekallikrein, and Factor XI on a negatively charged surface. Although prekallikrein and Factor XI circulate as a complex with HMW-kininogen, no physical association has been demonstrated between Factor XII and HMW-kininogen, nor has the order of adsorption to surfaces of these proteins been fully clarified. In this report, we explore the requirements for adsorption of HMWkininogen to a clot-promoting surface (kaolin), in purified systems, as well as in normal plasma and plasma genetically deficient in each ofthe proteins of the contact system. The fraction ofeach coagulant protein associated with the kaolin pellet was determined by measuring the difference in coagulant activity between the initial sample and supernatants after incubation with kaolin, or by directly quantifying the amount of '25I-HMW-kininogen that was associated with the kaolin pellet.In normal plasma, the adsorption of HMW-kininogen to kaolin increased as the quantity of kaolin was increased in the incubation mixture. However, the HMW-kininogen in Factor XII-deficient plasma did not absorb appreciably to kaolin. Furthermore, the quantity of HMW-kininogen Portions ofthis work were presented at the 55th session ofthe American Heart Association, November 15-18, 1982, Dallas, TX and were published in abstract form (1982, Circulation, 66:295 from prekallikrein-deficient plasma that adsorbed to kaolin was decreased as compared with normal plasma. These observations suggested that HMW-kininogen in plasma must be altered by a reaction involving both Factor XII and prekallikrein in order for HMW-kininogen to adsorb to kaolin, and to express its coagulant activity. Subsequently, the consequence of the inability of HMWkininogen to associate with a negatively charged surface results in decreased surface activation. This assessment was derived from the further observation of the lack of prekallikrein adsorption and the diminished Factor XI adsorption in both Factor XII-deficient and HMW-kininogen-deficient plasmas, since these two zymogens (prekallikrein and Factor XI) are transported to a negatively charged surface in complex with HMW-kininogen. The percentage of HMW-kininogen coagulant activity that adsorbed to kaolin closely correlated (r = 0.98, slope = 0.97) with the amount of 125I-HMW-kininogen adsorbed, suggesting that adsorption of HMW-kininogen results in the expression of its coagulant activity.Since kallikrein, which is known to cleave HMWkininogen, is generated when kaolin is added to plasma, we tested the hypothesis that proteolysis by kallikrein was responsible for the enhanced adsorption of HMW-kininogen to kaolin. When purified HMW-kininogen was incubated with purified kallikrein, its ability to adsorb to kaolin increased with time of digestion until a maximum was reached. Moreover, '25I-HMW-kininogen, after cleavage by kallikrein, had markedly increased affinity for kaolin than the unclea...

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“…Ainda em menor abundância em plaquetas ativadas, identificamos a proteína do complemento C3 e a proteína básica de plaqueta, que estão envolvidos em processos inflamatórios, e a fibronectina, uma proteína adesiva envolvida na formação do agregado plaquetário fazendo a ligação plaqueta-plaqueta através da O cininogênio, também identificado em menor abundância somente no proteoma do sedimento de plaquetas ativadas por trombina, tem um importante papel no início da coagulação sanguínea juntamente com o fator XII na ativação da tromboplastina plasmática (Saito, 1977). Além disso, a calicreína, ativada pelo fator XII, cliva o cininogênio liberando os peptídeos bradicinina ou kalidina, conhecidos como cininas vasoativas (Rocha e Silva, 1962), que tem ação sobre o tônus vascular.…”

Section: Sedimento De Plaquetasunclassified

Caracterização proteômica comparativa da agregação plaquetária induzida pela trombina e pela PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca

Oliveira¹

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Aos pesquisadores, pós-graduandos, estagiários e funcionários do LETA. Plaquetas são fragmentos celulares anucleados, derivados de megacariócitos, que estão envolvidos em diversos processos fisiológicos e patológicos, como coagulação, inflamação, trombose, aterosclerose, e metástase e angiogênese tumorais. Para executar estas funções, plaquetas ativadas secretam uma fração solúvel de moléculas presentes em seus conteúdos granulares, que passam a interagir com outras moléculas e células adjacentes ao local da injúria, e com os próprios receptores plaquetários. No entanto, os mecanismos que regem a secreção em plaquetas ainda são pouco conhecidos. Neste sentido, o objetivo deste estudo foi analisar comparativamente a agregação de plaquetas ativadas por dois diferentes agonistas enzimáticos: a trombina, um dos mais importantes agonistas plaquetários, e a PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca, que assim como a trombina, ativa plaquetas através dos receptores PAR-1 e PAR-4. Neste estudo foram utilizadas abordagens de espectrometria de massas e de bioinformática para caracterizar alterações nos proteomas do sedimento de plaquetas não ativadas e ativadas, e também, para em paralelo analisar as frações proteicas e peptídicas presentes no sobrenadante (secretoma). Nas análises do sedimento de plaquetas ativadas tanto por PA-BJ quanto por trombina, foi verificada a menor abundância das proteínas PBP, PF4, proteína S, fibronectina, fator V e alfa-1 antitripsina, entre outras, e que também foram identificadas no sobrenadante (secretadas), e o aumento de abundância das proteínas ADAM-10, tromboxano A2 sintase, integrina αIIb, miosina-9 e fosforilase b, que estão diretamente envolvidas na ativação/agregação. Por outro lado, verificamos que na secreção plaquetária induzida por trombina ocorreu o aumento de abundância de proteínas envolvidas na regulação da formação do coágulo, como a proteína S, PAI1 e antitrombina III, sugerindo que nos eventos disparados pela trombina, exista uma regulação rigorosa de sua ação no local da injúria vascular. Já na secreção induzida por PA-BJ, verificamos o aumento significativo das proteínas amiloide beta A4 e do fibrinogênio, envolvidas na ativação/agregação plaquetária, além da liberação e ativação de MMP1, indicando que esta metaloproteinase atue sinergicamente com a PA-BJ para a formação e estabilização do agregado plaquetário. Nas análises do secretoma de plaquetas não ativadas, identificamos pela primeira vez, a presença das proteínas catalase, anidrase carbônica, inibidor de elastase leucocitária e a glicoproteína rica em histidina, que estão envolvidas na inibição e regulação da ativação plaquetária. A análise da fração peptídica do sobrenadante plaquetário permitiu avaliar pela primeira vez o degradoma gerado no processo de agregação por PA-BJ e trombina. Ao longo desses dez anos de trabalho e aprendizado no Instituto Butantan eu tenho muito que agradecer! Obrigada a todos!!! "É necessário abrir os olhos e perceber que as coisas boas estão dentro de nós, onde os s...

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Purification of High Molecular Weight Kininogen and the Role of This Agent in Blood Coagulation

Abstract: A B S T R A C T Recent studies of individuals with high molecular weight (HMW)

Cited by 56 publications

References 29 publications

Interactions among Hageman factor, plasma prekallikrein, high molecular weight kininogen, and plasma thromboplastin antecedent.

Interactions among Hageman factor, plasma prekallikrein, high molecular weight kininogen, and plasma thromboplastin antecedent.

Cleavage of human high molecular weight kininogen markedly enhances its coagulant activity. Evidence that this molecule exists as a procofactor.

Caracterização proteômica comparativa da agregação plaquetária induzida pela trombina e pela PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca

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