Role of Bovine HMW Kininogen in Contact-Mediated Activation of Factor XII: Demonstration of a Nicked Form, “Active Kininogen”, With Maximal Cofactor Activity by Limited Proteolysis

1979 ◽  
Author(s):  
H. Kato ◽  
T. Sugo ◽  
N. Ikari ◽  
N. Hashimoto ◽  
S. Iwanaga ◽  
...  
Keyword(s):  
Light Chain ◽  
Positive Feedback ◽  
Limited Proteolysis ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Free Protein ◽  
Functional Regions ◽  
Cofactor Activity ◽  
Carboxyl Terminal

Bovine HMW kininogen (HMW-K) has a mol. wt. of 76,000 and the carboxyl-terminal 243 residues consisting of kinin, fragment 1.2 and light chain regions have been sequenced. The purpose of this study is to elucidate the functional regions of the kininogen which are required for the kaolin-mediated activation of Factor XII (XII) in the presence of prekallikrein (Prek). The results were as follows: (1) Kaolin-mediated activation of XII was accelerated 180 fold by adding optimum amounts of HMW-K. (2) The accelerating effect of HMW-K markedly increased by the brief treatment with kallikrein, indicating that a nicked HMW-K, named “Active Kininogen”, is most effective. (3) The same accelerating effect as HMW-K was observed with fragment 1.2-light chain. (4) Prek formed a complex with HMW-K and kinin-fragment 1.2-free protein. Kallikrein also formed a complex with kinin-fragment 1.2-free protein. (5) The adsorption of HMW-K on kaolin was inhibited by fragment 1.2. These results indicate that HMW-K accelerates kaolin-mediated activation or XII by forming a complex with Prek through the light chain region and by interacting with kaolin through the fragment 1.2 region. It was also suggested that plasma kallikrein plays an important role not only In the positive feedback activation of XII but also In the transformation of HMW-K to an “Active Kininogen”.

Role Of Hmw Kininogen In Polysaccharide Sulfate-Mediated Activation Of Factor XII

1981 ◽  
Author(s):  
H Kato ◽  
T Shimada ◽  
T Sugo ◽  
S Iwanaga
Keyword(s):  
Sulfur Content ◽  
Light Chain ◽  
Dextran Sulfate ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Charged Surfaces ◽  
Fluorogenic Peptide Substrate ◽  
Derivatives Of ◽  
Fluorogenic Peptide

We have shown that bovine HMW kininogen (HMW KGN) accelerates the activation of Factor XII (XII) and prekallikrein (Prek) in the presence of kaolin, adsorbing on kaolin through fragment 1.2 region and forming a complex with Prek through light chain region. The present study was undertaken to examine the functional role of HMW KGN in the activation of XII and Prek with other negatively-charged surfaces than kaolin. XII was incubated with a variety of substances, HMW KGN and Prek, and plasma kallikrein generated was estimated using a fluorogenic peptide substrate, Z- Phe-Arg-MCA.Among 8 kinds of polysaccharide sulfates (dextran sulfate, xylan sulfate, amylopectin sulfate, chondroitin polysulfate, heparan sulfate, amylose sulfate, heparin and carrageenin), amylose sulfate had the most potent ability to activate XII. The experiments using fourteen amylose sulfates with different sulfur content and different viscosity revealed that some amylose sulfates with a higher sulfur content (18%) activated strikingly XII and Prek in the presence of HMW KGN. In the reaction, fragment 1.2-light chain region in HMW KGN was shown to be essential, using the various derivatives of HMW KGN. The rate of XII activation was quite dependent on the concentration of HMW KGN and amylose sulfate. At higher concentrations, both of them inhibited the activation reaction. On the other hand, sulfatide has been shown to accelerate the activation of XII with plasma kallikrein by Fujikawa et al. and the reaction was inhibited by the addition of HMW KGN. In the present study, the activation of XII and Prek with sulfatides as estimated by the generation of kallikrein, was accelerated by HMW KGN. This reaction was also dependent on the concentrations of sulfatides and HMW KGN, and fragment 1.2-light chain region in HMW KGN was shown to be essential.These results indicate that HMW KGN is required to accelerate the activation of XII and Prek with polysaccharide sulfate and sulfatide as well as kaolin.

scholarly journals The Carboxyl-Terminal Domain of TraR, a Streptomyces HutC Family Repressor, Functions in Oligomerization

2008 ◽  
Vol 190 (21) ◽  
pp. 7164-7169 ◽  
Author(s):  
Masakazu Kataoka ◽  
Takeshi Tanaka ◽  
Toshiyuki Kohno ◽  
Yusuke Kajiyama
Keyword(s):  
Limited Proteolysis ◽  
Sequence Information ◽  
Structural Domains ◽  
Terminal Domain ◽  
Tetramer Formation ◽  
Regulated Expression ◽  
Carboxyl Terminal ◽  
Two Hybrid

ABSTRACT Efficient conjugative transfer of the Streptomyces plasmid pSN22 is accomplished by regulated expression of the tra operon genes, traA, traB, and spdB. The TraR protein is the central transcriptional repressor regulating the expression of the tra operon and itself and is classified as a member of the HutC subfamily in the helix-turn-helix (HTH) GntR protein family. Sequence information predicts that the N-terminal domain (NTD) of TraR, containing an HTH motif, functions in binding of DNA to the cis element; however, the function of the C-terminal region remains obscure, like that for many other GntR family proteins. Here we demonstrate the domain structure of the TraR protein and explain the role of the C-terminal domain (CTD). The TraR protein can be divided into two structural domains, the NTD of M1 to R95 and the CTD of Y96 to E246, revealed by limited proteolysis. Domain expression experiments revealed that both domains retained their function. An in vitro pull-down assay using recombinant TraR proteins revealed that TraR oligomerization depended on the CTD. A bacterial two-hybrid system interaction assay revealed that the minimum region necessary for this binding is R95 to P151. A mutant TraR protein in which Leu121 was replaced by His exhibited a loss of both oligomerization ability and repressor function. An in vitro cross-linking assay revealed preferential tetramer formation by TraR and the minimum CTD. These results indicate that the C-terminal R95-to-P151 region of TraR functions to form an oligomer, preferentially a tetramer, that is essential for the repressor function of TraR.

scholarly journals Effect of cleavage of the heavy chain of human plasma kallikrein on its functional properties

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 311-318 ◽  
Author(s):  
RW Colman ◽  
YT Wachtfogel ◽  
U Kucich ◽  
G Weinbaum ◽  
S Hahn ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Heavy Chain ◽  
Light Chain ◽  
Human Neutrophils ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
C1 Inhibitor ◽  
Amidolytic Activity ◽  
Coagulant Activity

Abstract Human plasma kallikrein consists of an N-terminal heavy chain of molecular weight (mol wt) 52,000, linked by disulfide bonds to two light chain variants (mol wt 36,000 or 33,000). Although the active catalytic site of kallikrein resides on the C-terminal light chain, the role of the N-terminal heavy chain is less clear. We therefore studied an enzyme designated beta-kallikrein, containing a single cleavage in the heavy chain (mol wt 28,000 + 18,000) and compared it to the enzyme, alpha-kallikrein, with an intact heavy chain. The rates of inactivation by C1 inhibitor of plasma alpha- and beta-kallikreins were kinetically identical, as measured by residual amidolytic activity, after various times of incubation with the inhibitor. Both enzymes reacted completely with C1 inhibitor after 18 hours and formed identical C1 inhibitor- kallikrein complexes of mol wt 195,000. The rate of activation of factor XII by alpha-kallikrein and beta-kallikrein was similar. In contrast, the rate of cleavage of high molecular weight kininogen (HMWK) by alpha-kallikrein was at least fivefold faster and the ratio of coagulant activity to amidolytic activity was fourfold greater than for beta-kallikrein. Plasma alpha-kallikrein, at concentrations potentially achievable in plasma, induced aggregation of neutrophils, but beta-kallikrein failed to elicit this response. In addition, human neutrophils pretreated with cytochalasin B released 2.46 +/- 0.10 microgram/10(7) cells of elastase antigen, but beta-kallikrein released only 0.25 +/- 0.10 micrograms/10(7) cells. These observations suggest that cleavage of the heavy chain influences the rate of cleavage of HMWK and decreases its coagulant activity. Moreover, an intact heavy chain appears to be requisite to support the ability of kallikrein to aggregate neutrophils and release elastase.

scholarly journals Role of plasma kallikrein in diabetes and metabolism

2013 ◽  
Vol 110 (09) ◽  
pp. 434-441 ◽  
Author(s):  
Qunfang Zhou ◽  
Ward Fickweiler ◽  
Edward P. Feener
Keyword(s):  
Vascular Function ◽  
Physiological Role ◽  
Coagulation Factor ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Pressure Regulation ◽  
Pharmacological Studies ◽  
Coagulation Pathway ◽  
Insight Into

SummaryPlasma kallikrein (PK) is a serine protease generated from plasma prekallikrein, an abundant circulating zymogen expressed by the Klkb1 gene. The physiological actions of PK have been primarily attributed to its production of bradykinin and activation of coagulation factor XII, which promotes inflammation and the intrinsic coagulation pathway. Recent genetic, molecular, and pharmacological studies of PK have provided further insight into its role in physiology and disease. Genetic analyses have revealed common Klkb1 variants that are association with blood metabolite levels, hypertension, and coagulation. Characterisation of animal models with Klkb1 deficiency and PK inhibition have demonstrated effects on inflammation, vascular function, blood pressure regulation, thrombosis, haemostasis, and metabolism. These reports have also identified a host of PK substrates and interactions, which suggest an expanded physiological role for this protease beyond the bradykinin system and coagulation. The review summarises the mechanisms that contribute to PK activation and its emerging role in diabetes and metabolism.

Structure - Function Relationships of Human High MW Kininogen(HMWK), A Cofactor in Contact Activation of Human Plasma

1979 ◽  
Author(s):  
D. M. Kerbiriou ◽  
J.H. Griffin
Keyword(s):  
Human Plasma ◽  
Light Chain ◽  
Procoagulant Activity ◽  
Free Form ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Free Molecule ◽  
Contact Activation ◽  
Single Polypeptide Chain ◽  
Single Polypeptide

HMWK has been isolated from human plasma as a single polypeptide chain of apparent 110,000 MW on SDS gels. Purified plasma kallikrein cleaved HMWK to produce kinin and a kinin-free molecule made up of two disulfide-linked polypeptide chains with apparent MW of 65,000 and 44,000. Following reduction and carboxymethylation of the two-chain, kinin-free molecule, a histidine-rich light chain was isolated that quantitatively retained the full procoagulant activity of the native molecule. HMWK was digested with a bacterial protease to produce kinin and polypeptides that have no procoagulant activity. From this digest, cationic fragments were isolated which possess a high histidine (36%) and glycine (36%) content that is similar to the composition of Fragment 1.2 cleaved from bovine HMWK by bovine plasma kallikrein. When kaolin was added to normal human plasma containing 125I-HMWK and 131I-Factor XII , the HMWK was rapidly cleaved with a half-time of cleavage of 40 ± 10 sec which was similar to the kinetics of cleavage of Factor XII. This surface-initiated cleavage of 125I-HMWK in plasma did not occur in plasmas deficient in either prekallikrein or Factor XII, but occurred when purified kallikrein was added. These observations suggest that when kaolin is added to plasma, kallikrein cleaved HMWK to release kinin and to give rise to a two-chain, kinin-free form of HMWK whose ability to correct the clotting deficiency of HMWK-deficient plasma resides totally in the histidine-rich light chain of the molecule.

scholarly journals Effect of cleavage of the heavy chain of human plasma kallikrein on its functional properties

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 311-318 ◽  
Author(s):  
RW Colman ◽  
YT Wachtfogel ◽  
U Kucich ◽  
G Weinbaum ◽  
S Hahn ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Heavy Chain ◽  
Light Chain ◽  
Human Neutrophils ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
C1 Inhibitor ◽  
Amidolytic Activity ◽  
Coagulant Activity

Human plasma kallikrein consists of an N-terminal heavy chain of molecular weight (mol wt) 52,000, linked by disulfide bonds to two light chain variants (mol wt 36,000 or 33,000). Although the active catalytic site of kallikrein resides on the C-terminal light chain, the role of the N-terminal heavy chain is less clear. We therefore studied an enzyme designated beta-kallikrein, containing a single cleavage in the heavy chain (mol wt 28,000 + 18,000) and compared it to the enzyme, alpha-kallikrein, with an intact heavy chain. The rates of inactivation by C1 inhibitor of plasma alpha- and beta-kallikreins were kinetically identical, as measured by residual amidolytic activity, after various times of incubation with the inhibitor. Both enzymes reacted completely with C1 inhibitor after 18 hours and formed identical C1 inhibitor- kallikrein complexes of mol wt 195,000. The rate of activation of factor XII by alpha-kallikrein and beta-kallikrein was similar. In contrast, the rate of cleavage of high molecular weight kininogen (HMWK) by alpha-kallikrein was at least fivefold faster and the ratio of coagulant activity to amidolytic activity was fourfold greater than for beta-kallikrein. Plasma alpha-kallikrein, at concentrations potentially achievable in plasma, induced aggregation of neutrophils, but beta-kallikrein failed to elicit this response. In addition, human neutrophils pretreated with cytochalasin B released 2.46 +/- 0.10 microgram/10(7) cells of elastase antigen, but beta-kallikrein released only 0.25 +/- 0.10 micrograms/10(7) cells. These observations suggest that cleavage of the heavy chain influences the rate of cleavage of HMWK and decreases its coagulant activity. Moreover, an intact heavy chain appears to be requisite to support the ability of kallikrein to aggregate neutrophils and release elastase.

Cold Promoted Activation of Factor VII

1972 ◽  
Vol 28 (02) ◽  
pp. 169-181 ◽  
Author(s):  
H Gjønnæss
Keyword(s):  
Low Temperature ◽  
Oral Contraceptives ◽  
Fold Increase ◽  
Factor Vii ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Overnight Incubation ◽  
Esterolytic Activity

SummaryThe activating principle (CPA) of the factor VII activation seen in plasmas of women taking oral contraceptives after overnight incubation of the plasmas at 0° C was investigated. The reaction was dependent on low temperature, and factor XII was indispensable. Concomitant with the activation of factor VII a 10–30 fold increase in TAME esterolytic activity was observed together with a near 100 per cent drop in plasma kininogen concentration. The results indicated that the activation of factor VII is linked to activation of the kallikrein system, and that the activator may be plasma kallikrein.

Functional Characterization of a Variant Factor XII (F XII Locarno) in a Cross Reacting Material Positive F XII Deficient Plasma

1992 ◽  
Vol 67 (02) ◽  
pp. 219-225 ◽  
Author(s):  
Walter A Wuillemin ◽  
Miha Furlan ◽  
Hans Stricker ◽  
Bernhard Lämmle
Keyword(s):  
Dextran Sulfate ◽  
Functional Characterization ◽  
Healthy Woman ◽  
Chain Molecule ◽  
Plasma Kallikrein ◽  
Factor Xii ◽  
Single Chain ◽  
Sulfate Adsorption ◽  
Prolonged Incubation ◽  
Functional Studies

SummaryThe plasma of a healthy woman was found to contain half normal factor XII (FXII) antigen level (0.46 U/ml) without any FXII clotting activity (<0.01 U/ml). The variant FXII in this plasma, denoted as FXII Locarno, was partially characterized by immunological and functional studies on the proposita’s plasma. FXII Locarno is a single chain molecule with the same size (M r = 80 kDa) as normal FXII. Isoelectric focusing suggested an excess of negative charge in the variant FXII as compared to normal FXII. In contrast to FXII in normal plasma, FXII Locarno was not proteolytically cleaved upon prolonged incubation of proposita’s plasma with dextran sulfate. Adsorption to kaolin was similar for both, abnormal and normal FXII. Incubation of the proposita’s plasma with dextran sulfate and exogenous plasma kallikrein showed normal cleavage of FXII Locarno outside of the tentative disulfide loop Cys340-Cys467, but only partial cleavage within this disulfide loop. Furthermore, plasma kallikrein-cleaved abnormal FXII showed neither amidolytic activity nor proteolytic activity against factor XI and plasma prekallikrein.These results suggest a structural alteration of FXII Locarno, affecting the plasma kallikrein cleavage site Arg353-Val354 and thus formation of activated FXII (a-FXIIa).

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