scholarly journals Relationship between structure and correcting activity of bovine high molecular weight kininogen upon the clotting time of fitzgerald-trait plasma

1979 ◽  
Vol 149 (4) ◽  
pp. 847-855 ◽  
Author(s):  
AG Scicli ◽  
R Waldmann ◽  
JA Guimaraes ◽  
G Scicli ◽  
OA Carretero ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
Plasma Kallikrein ◽  
High Molecular Weight Kininogen ◽  
Clotting Time ◽  
Active Inhibitor ◽  
Charged Surfaces ◽  
Bovine Plasma

Bovine high molecular weight kininogen (bHMWK) partially corrects the activated plasma thromboplastin time (aPTT) of Fitzgerald trait plasma which is congenitally deficient in HMWK. The relationship between the structure and activity of HMWK was clarified by studying the effects of different fragments of bHMWK on the aPTT of Fitzgerald-trait plasma. The peptides studied were lys-bradykinin-free HMWK, bradykinin-fragment 1-2-free HMWK, heavy chain, fragment 1-2-light chain, and light chain. All fragments were tested in equimolar concentrations. Bradykinin-fragment 1-2-free HMWK, heavy chain, and light chain have little or no correcting activity upon Fitzgerald-trait plasma aPTr. Fragment 1-2 light chain has the same correcting activity as intact bHMWK, while that of lys-bradykinin-free HMWK appears to be higher. Both fragment 1-2 and fragment 2 inhibit the clotting time of normal human plasma. When compared on a molar basis, fragment 2 is a more active inhibitor than fragment 1-2. When the effects of bovine plasma kallikrein upon bHMWK and hHMWK were studied, it was found that it released kinins from both kininogens. However, while the correcting activity of bHMWK was completely destroyed after 60 min of incubation, that of hHMWK was fully retained. These data suggest that: (a) the active part of bHMWK is comprised of the fragment 1-2 light chain portion; (b) fragment 1-2 or fragment 2 is the binding site to negatively charged surfaces, while the light chain interacts with other components of the surface-mediated reactions; and (c) bovine plasma kallikrein releases kinins, but probably does not cause the release of fragment 1-2 from human HMWK.

Corrections - High Molecular Weight Kininogen or its Light Chain Protects Human Plasma Kallikrein from Inactivation by Plasma Protease Inhibitors

Biochemistry ◽  
1982 ◽  
Vol 21 (12) ◽  
pp. 3036-3036
Author(s):  
Marc Schapira ◽  
Cheryl Scott ◽  
Ann James ◽  
Lee Silver ◽  
Frederich Kueppers ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
High Molecular Weight ◽  
Protease Inhibitors ◽  
Light Chain ◽  
Plasma Kallikrein ◽  
High Molecular Weight Kininogen

High molecular weight kininogen or its light chain protects human plasma kallikrein from inactivation by plasma protease inhibitors

Biochemistry ◽  
1982 ◽  
Vol 21 (3) ◽  
pp. 567-572 ◽  
Author(s):  
Marc Schapira ◽  
Cheryl F. Scott ◽  
Ann James ◽  
Lee D. Silver ◽  
Friedrich Kueppers ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
High Molecular Weight ◽  
Protease Inhibitors ◽  
Light Chain ◽  
Plasma Kallikrein ◽  
High Molecular Weight Kininogen

scholarly journals High molecular weight kininogen binds to platelets by its heavy and light chains and when bound has altered susceptibility to kallikrein cleavage

Blood ◽  
1992 ◽  
Vol 79 (5) ◽  
pp. 1233-1244 ◽  
Author(s):  
FJ Meloni ◽  
EJ Gustafson ◽  
AH Schmaier
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
Competitive Inhibitor ◽  
Light Chains ◽  
High Molecular Weight Kininogen ◽  
Cell Binding ◽  
Platelet Surface ◽  
Platelet Binding

Abstract The unstimulated platelet surface contains a specific and saturable binding site for high molecular weight kininogen (HK) and low molecular weight kininogen (LK). Investigations were performed with purified heavy and light chains of HK to determine which portion(s) of the HK molecule binds to the platelet surface. Purified 64-Kd heavy chain of HK and 56-Kd light chain of HK, independently, inhibited 125I-HK binding to unstimulated platelets with a 50% inhibitory concentration (IC50) of 84 nmol/L (apparent Ki, 30 nmol/L) and 30 nmol/L (apparent Ki, 11 nM), respectively. The ability of each of the purified chains of HK to independently inhibit 125I-HK binding was not due to cleavage, reduction, and alkylation of the protein, because two-chain HK, produced by treating HK the same way as purifying the separate chains, inhibited binding similarly to intact HK. Further, purified LK alone inhibited 125I-HK binding to platelets (Ki, 17 +/- 1 nmol/L, n = 7). The 64-Kd heavy chain of HK was a competitive inhibitor on a reciprocal plot of 125I-HK-platelet binding with an apparent Ki of 28 +/- 6 nmol/L (n = 4). Independently, purified 56-Kd light chain of HK was also found to be a competitive inhibitor of 125I-HK-platelet binding, with an apparent Ki of 11 +/- 3 nmol/L (mean +/- SEM, n = 4). These indirect studies indicated that HK binds to platelets by two portions of the molecule, one on the heavy chain and another on the light chain. Studies with 125I-light chain of HK showed that it specifically bound directly to platelets in the presence of zinc, since it was blocked by HK, light chain of HK, or EDTA, but not by LK, C1s, C1 inhibitor, plasmin, factor XIII, or fibrinogen. Purified light chain of HK did not inhibit direct 125I-LK binding to platelets. HK was found to bind to platelets in an unmodified form. HK bound to platelets was cleaved by plasma or urinary kallikrein at a slower rate than the same concentration of soluble HK or HK bound and subsequently eluted from the platelet surface. Cleavage of platelet-bound HK correlated with bradykinin liberation. These studies indicate that HK has two domains on its molecule that bind to platelets. Further, platelet-bound HK is protected from kallikreins' proteolysis. This latter finding suggests that cell binding may modify the rate of bradykinin liberation from HK.

scholarly journals Characterization of human high molecular weight kininogen. Procoagulant activity associated with the light chain of kinin-free high molecular weight kininogen.

1978 ◽  
Vol 147 (2) ◽  
pp. 488-499 ◽  
Author(s):  
R E Thompson ◽  
R Mandle ◽  
A P Kaplan
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
Coagulation Factor ◽  
Antigenic Determinants ◽  
High Molecular Weight Kininogen ◽  
Single Chain ◽  
Apparent Loss ◽  
Coagulant Activity

Human high molecular weight (HMW) kininogen has been isolated and was found to be a single chain protein of approximately equal to 120,000 daltons. Upon digestion with plasma kallikrein bradykinin is generated, and SDS gel electrophoresis of the kinin-free protein reveals an apparent loss in size of 15,000 daltons. The kinin-free kininogen retains full activity as a coagulation factor and consists of two chains: a heavy chain of approximately equal to 66,000 daltons disulfide-linked to a light chain of 37,000 daltons. The heavy chain of HMW kininogen shares antigenic determinants with LMW kininogen and possesses no detectable coagulant activity. The isolated light chain is shown to be responsible for the coagulant activity of HMW kininogen and contains a unique antigenic determinant that distinguishes HMW kininogen from low molecular weight kininogen.

scholarly journals High molecular weight kininogen binds to platelets by its heavy and light chains and when bound has altered susceptibility to kallikrein cleavage

Blood ◽  
1992 ◽  
Vol 79 (5) ◽  
pp. 1233-1244 ◽  
Author(s):  
FJ Meloni ◽  
EJ Gustafson ◽  
AH Schmaier
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
Competitive Inhibitor ◽  
Light Chains ◽  
High Molecular Weight Kininogen ◽  
Cell Binding ◽  
Platelet Surface ◽  
Platelet Binding

The unstimulated platelet surface contains a specific and saturable binding site for high molecular weight kininogen (HK) and low molecular weight kininogen (LK). Investigations were performed with purified heavy and light chains of HK to determine which portion(s) of the HK molecule binds to the platelet surface. Purified 64-Kd heavy chain of HK and 56-Kd light chain of HK, independently, inhibited 125I-HK binding to unstimulated platelets with a 50% inhibitory concentration (IC50) of 84 nmol/L (apparent Ki, 30 nmol/L) and 30 nmol/L (apparent Ki, 11 nM), respectively. The ability of each of the purified chains of HK to independently inhibit 125I-HK binding was not due to cleavage, reduction, and alkylation of the protein, because two-chain HK, produced by treating HK the same way as purifying the separate chains, inhibited binding similarly to intact HK. Further, purified LK alone inhibited 125I-HK binding to platelets (Ki, 17 +/- 1 nmol/L, n = 7). The 64-Kd heavy chain of HK was a competitive inhibitor on a reciprocal plot of 125I-HK-platelet binding with an apparent Ki of 28 +/- 6 nmol/L (n = 4). Independently, purified 56-Kd light chain of HK was also found to be a competitive inhibitor of 125I-HK-platelet binding, with an apparent Ki of 11 +/- 3 nmol/L (mean +/- SEM, n = 4). These indirect studies indicated that HK binds to platelets by two portions of the molecule, one on the heavy chain and another on the light chain. Studies with 125I-light chain of HK showed that it specifically bound directly to platelets in the presence of zinc, since it was blocked by HK, light chain of HK, or EDTA, but not by LK, C1s, C1 inhibitor, plasmin, factor XIII, or fibrinogen. Purified light chain of HK did not inhibit direct 125I-LK binding to platelets. HK was found to bind to platelets in an unmodified form. HK bound to platelets was cleaved by plasma or urinary kallikrein at a slower rate than the same concentration of soluble HK or HK bound and subsequently eluted from the platelet surface. Cleavage of platelet-bound HK correlated with bradykinin liberation. These studies indicate that HK has two domains on its molecule that bind to platelets. Further, platelet-bound HK is protected from kallikreins' proteolysis. This latter finding suggests that cell binding may modify the rate of bradykinin liberation from HK.

scholarly journals High molecular weight kininogen binds to Mac-1 on neutrophils by its heavy chain (domain 3) and its light chain (domain 5).

1994 ◽  
Vol 269 (30) ◽  
pp. 19307-19312 ◽  
Author(s):  
Y.T. Wachtfogel ◽  
R.A. DeLa Cadena ◽  
S.P. Kunapuli ◽  
L. Rick ◽  
M. Miller ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
High Molecular Weight Kininogen ◽  
Domain 3

Cleaved High Molecular Weight Kininogen Induces Endothelial Progenitor Cell Senescence by Stimulation of JNK/FOXO4/MnSOD Pathway.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2181-2181
Author(s):  
Xuemei Zhu ◽  
Robert W. Colman ◽  
Yi Wu
Keyword(s):  
Molecular Weight ◽  
Transcription Factor ◽  
High Molecular Weight ◽  
Heavy Chain ◽  
Light Chain ◽  
Endothelial Progenitor Cell ◽  
Progenitor Cell ◽  
High Molecular Weight Kininogen ◽  
Endothelial Progenitor ◽  
Stimulation Of

Abstract Abstract 2181 Objectives: High molecular weight kininogen (HK) is a major component of plasma kallikrein-kinin system (KKS), and is cleaved to its active form (HKa) upon the activation of this system. Although the KKS activation is widely involved in a variety of pathological settings, the activities of its activation product HKa remain largely unknown. Recently we have reported that HKa accelerates the onset of endothelial progenitor cell (EPC) senescence by induction of reactive oxygen species (Arterioscler Thromb Vasc Biol. 2011;31:883), whereas the mechanism is not clear. In this study, we investigated whether HKa induces EPC senescence via stimulation of c-Jun N-terminal kinases (JNK)-related pathway. Methods: Human EPCs were treated with HKa in different concentrations for 72 hours, which were followed by the measurement of phospharylation of JNK at Thr183/tyr185 and transcription factor FOXO4 at Thr451, as well as expression of Mn-superoxide dismutase (MnSOD) at protein and mRNA level. To narrow down the functional domain of HKa, recombinant proteins of human HK heavy chain (HC, 19–380aa) and light chain (LC, 390–644aa) were generated for determining which domain(s) mediates the effect of HKa. Results: In a concentration-dependent manner, HKa treatment induced phosphorylation of JNK at Thr183/Tyr185, and its downstream phosphorylation of transcription factor FOXO4 at Thr451. Concomitantly, HKa upregulated the expression of MnSOD at protein and mRNA levels as measured by Western blot and real time RT-PCR. However, it did not affect the expression of catalase. Similar to HKa, the heavy chain (HC), but not the light chain, increased the percentage of senescent EPCs (63%±6.6 for HC v.s. 77.5%±6.02 for HKa). Moreover, HC at 100 nM increased FOXO4 phosphorylation at Thr451 and the expression of MnSOD in EPCs. Besides, both of HKa and its HC stimulated the production of intracellular H2O2. Conclusion: These above results demonstrate that HKa accelerates the onset of EPC senescence by stimulating JNK/FOXO4/MnSOD pathway, and its effect is mediated by the HC. Disclosures: No relevant conflicts of interest to declare.

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