23 results on '"Kaplan AP"'
Search Results
2. Protease activity in single-chain prekallikrein.
- Author
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Ivanov I, Verhamme IM, Sun MF, Mohammed B, Cheng Q, Matafonov A, Dickeson SK, Joseph K, Kaplan AP, and Gailani D
- Subjects
- Amino Acid Substitution, Animals, Factor XII metabolism, HEK293 Cells, Humans, Kininogen, High-Molecular-Weight metabolism, Mice, Inbred C57BL, Prekallikrein chemistry, Prekallikrein genetics, Proteolysis, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Blood Coagulation, Bradykinin metabolism, Prekallikrein metabolism
- Abstract
Prekallikrein (PK) is the precursor of the trypsin-like plasma protease kallikrein (PKa), which cleaves kininogens to release bradykinin and converts the protease precursor factor XII (FXII) to the enzyme FXIIa. PK and FXII undergo reciprocal conversion to their active forms (PKa and FXIIa) by a process that is accelerated by a variety of biological and artificial surfaces. The surface-mediated process is referred to as contact activation. Previously, we showed that FXII expresses a low level of proteolytic activity (independently of FXIIa) that may initiate reciprocal activation with PK. The current study was undertaken to determine whether PK expresses similar activity. Recombinant PK that cannot be converted to PKa was prepared by replacing Arg371 with alanine at the activation cleavage site (PK-R371A, or single-chain PK). Despite being constrained to the single-chain precursor form, PK-R371A cleaves high-molecular-weight kininogen (HK) to release bradykinin with a catalytic efficiency ∼1500-fold lower than that of kallikrein cleavage of HK. In the presence of a surface, PK-R371A converts FXII to FXIIa with a specific activity ∼4 orders of magnitude lower than for PKa cleavage of FXII. These results support the notion that activity intrinsic to PK and FXII can initiate reciprocal activation of FXII and PK in solution or on a surface. The findings are consistent with the hypothesis that the putative zymogens of many trypsin-like proteases are actually active proteases, explaining their capacity to undergo processes such as autoactivation and to initiate enzyme cascades., (© 2020 by The American Society of Hematology.)
- Published
- 2020
- Full Text
- View/download PDF
3. Nomenclature of factor XI and the contact system.
- Author
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Schmaier AH, Emsley J, Feener EP, Gailani D, Govers-Riemslag JWP, Kaplan AP, Maas C, Morrissey JH, Renné T, Sidelmann JJ, and Meijers JCM
- Subjects
- Consensus, Factor XI chemistry, Factor XII chemistry, Factor XII classification, Factor XII metabolism, Humans, Kininogen, High-Molecular-Weight chemistry, Kininogen, High-Molecular-Weight classification, Kininogen, High-Molecular-Weight metabolism, Molecular Structure, Prekallikrein chemistry, Prekallikrein classification, Prekallikrein metabolism, Structure-Activity Relationship, Blood Coagulation, Factor XI classification, Factor XI metabolism, Terminology as Topic
- Published
- 2019
- Full Text
- View/download PDF
4. Assessment of the role of heparan sulfate in high molecular weight kininogen binding to human umbilical vein endothelial cells.
- Author
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Fernando LP, Fernando AN, Joseph K, and Kaplan AP
- Subjects
- Antibodies pharmacology, Cells, Cultured, Endothelium, Vascular metabolism, Heparin Lyase pharmacology, Humans, Kallikreins, Keratins physiology, Kinetics, Membrane Glycoproteins physiology, Protein Binding, Receptors, Complement physiology, Umbilical Veins, Zinc, Endothelium, Vascular cytology, Heparitin Sulfate physiology, Kininogen, High-Molecular-Weight metabolism
- Abstract
The assembly and activation of the kinin forming system components on human umbilical vein endothelial cells (HUVEC) have been studied in great detail. Proteins such as gC1qR, cytokeratin-1 and u-PAR have been identified to be responsible for Zn2+-dependent binding of high molecular weight kininogen (HK) to HUVEC. Heparan sulfate has also been shown to have a major role in Zn2+-dependent binding of HK to the endothelial cell line, Ea.hy 926. In this study, we have analyzed the possible contribution of heparan sulfate to high molecular weight kininogen binding to HUVEC using multiple approaches. The presence of heparan sulfate on HUVEC was analyzed by staining with an antibody specific for heparan sulfate. Incubation of the cells with bacterial heparinases removed the heparan sulfate from the cell surface to the level seen with a control antibody, however, the Zn2+-dependent binding of HK was not affected. Further, blocking of heparan sulfate with a specific antibody to heparan sulfate even after digestion with heparinases did not reduce HK binding whereas antibodies to the proteins gC1qR and cytokeratin-1 consistently reduced the binding of HK to the endothelial cells. The binding intensities of FITC-labeled HK were similar in heparinase-treated and -untreated HUVEC. The rate of kallikrein formation by the assembly of factor XII, HK and PK were similar in both heparinase-treated and non-treated HUVEC. All of these data indicate that heparan sulfate does not contribute significantly to HK binding to HUVEC.
- Published
- 2003
- Full Text
- View/download PDF
5. Intrinsic coagulation, thrombosis, and bleeding.
- Author
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Kaplan AP
- Subjects
- Enzyme Activation, Factor XI Deficiency blood, Factor XII Deficiency blood, Hemorrhagic Disorders blood, Kininogens deficiency, Plasma metabolism, Prekallikrein deficiency, Blood Coagulation physiology, Factor IX physiology, Hemorrhagic Disorders etiology, Thrombin metabolism, Thromboplastin metabolism, Thrombosis blood
- Published
- 1996
6. Human high molecular weight kininogen binds to human umbilical vein endothelial cells via its heavy and light chains.
- Author
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Reddigari SR, Kuna P, Miragliotta G, Shibayama Y, Nishikawa K, and Kaplan AP
- Subjects
- Binding Sites, Cells, Cultured, Humans, Kininogens chemistry, Molecular Weight, Umbilical Veins, Zinc pharmacology, Endothelium, Vascular metabolism, Kininogens metabolism
- Abstract
High molecular weight kininogen (HK) is a multifunctional plasma glycoprotein that occupies a critical position in pathways that link inflammation and coagulation. Excision of the vasoactive peptide bradykinin by plasma kallikrein results in kinin-free HK that consists of a 65-Kd N-terminal heavy chain (HK-HC) linked to the C-terminal 45-Kd light chain (HK-LC) by a disulfide bridge. HK-HC is an inhibitor of SH-proteases and HK-LC contains the binding sites for coagulation cofactors prekallikrein and factor XI. HK has previously been shown to bind specifically to human umbilical vein endothelial cells (HUVEC) in a zinc(2+)-dependent manner by a single class of high-affinity binding sites. We have further characterized that interaction in order to determine the cell-binding regions of HK. Competition binding experiments have indicated that either HK-LC or HK-HC was able to inhibit the binding of labeled HK with a 50% inhibitory concentration (IC50) of 77 nmol/L and 89 nmol/L, respectively. Cleaved two-chain HK (HKa) had an IC50 of 73 nmol/L, whereas uncleaved HK had an IC50 of 335 nmol/L. Direct binding experiments have indicated that HUVEC bind both purified [125I]HK-HC and [125I]HK-LC in a zinc(2+)-dependent manner and that HK-LC did not displace bound HK-HC. The light chain of low molecular weight kininogen or prekallikrein-binding region of HK did not inhibit the binding of HK to HUVEC. Our results, therefore, indicate that (1) HK is capable of binding to endothelial cells via both heavy and light chain moieties, (2) HKa has a higher affinity to HUVEC, and (3) purified heavy and light chains are capable of directly binding to HUVEC. The data are consistent with the presence of a single high-affinity site for HK on endothelial cells within which are subsites that bind to heavy and light chains.
- Published
- 1993
7. Activation of factor XI in plasma is dependent on factor XII.
- Author
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Brunnée T, La Porta C, Reddigari SR, Salerno VM, Kaplan AP, and Silverberg M
- Subjects
- Autoradiography, Blood Coagulation, Electrophoresis, Polyacrylamide Gel, Factor XI isolation & purification, Factor XI Deficiency blood, Factor XII isolation & purification, Factor XII Deficiency blood, Humans, Iodine Radioisotopes, Kinetics, Macromolecular Substances, Molecular Weight, Thrombin metabolism, Factor XI metabolism, Factor XII metabolism, Factor XIa metabolism
- Abstract
The activation of factor XI initiates the intrinsic coagulation pathway. Until recently it was believed that the main activator of factor XI is factor XIIa in conjunction with the cofactor high molecular weight kininogen on a negatively charged surface. Two recent reports have presented evidence that in a purified system factor XI is activatable by thrombin together with the soluble polyanion dextran sulfate. To assess the physiological relevance of these findings we studied the activation of factor XI in normal and factor XII-deficient plasma. We used either kaolin/cephalin or dextran sulfate as a surface for the intrinsic coagulation pathway, tissue factor to generate thrombin via the extrinsic pathway, or the addition of alpha-thrombin directly. 125I-factor XI, added to factor XI-deficient plasma at physiologic concentrations (35 nmol/L), is rapidly cleaved on incubation with kaolin. The kinetics appear to be exponential with half the maximum cleavage at 5 minutes. Similar kinetics of factor XI cleavage are seen when 40 nmol/L factor XIIa (equal to 10% of factor XII activation) is added to factor XII-deficient plasma if an activating surface is provided. Tissue factor (1:500) added to plasma did not induce cleavage of factor XI during a 90-minute incubation, although fibrin formation within 30 seconds indicated that thrombin was generated via the extrinsic pathway. Adding 1 mumol/L alpha-thrombin (equivalent to 50% prothrombin activation) directly to factor XII deficient or normal plasma (with or without kaolin/cephalin/Ca2+ or dextran sulfate) led to instantaneous fibrinogen cleavage, but again no cleavage of factor XI was observable. We conclude that in plasma surroundings factor XI is not activated by thrombin, and that proposals of thrombin initiation of the intrinsic coagulation cascade are not supportable.
- Published
- 1993
8. Generation of vasoactive peptide bradykinin from human umbilical vein endothelium-bound high molecular weight kininogen by plasma kallikrein.
- Author
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Nishikawa K, Shibayama Y, Kuna P, Calcaterra E, Kaplan AP, and Reddigari SR
- Subjects
- Amino Acid Sequence, Cells, Cultured, Humans, Immunoenzyme Techniques, Kinetics, Kinins, Molecular Sequence Data, Umbilical Veins, Bradykinin metabolism, Endothelium, Vascular metabolism, Kallikreins metabolism, Kininogens metabolism, Protein Precursors metabolism
- Abstract
High molecular weight kininogen (HK) is a multifunctional plasma glycoprotein that occupies a critical position in pathways that link inflammation and coagulation. It is an inhibitor of sulfhydryl proteases and has procoagulant properties. It is also a source of the vasoactive peptide bradykinin (BK). It has been previously shown that HK binds to human umbilical vein endothelial cells (HUVEC) in culture. We have further characterized that interaction herein. Immunohistochemical experiments have indicated that when freshly obtained umbilical vein segments were treated with HK, washed, and probed with anti-HK antibodies, HK was localized on the endothelium. We next determined whether HUVEC-bound HK can be cleaved by plasma kallikrein to release BK. Cultured HUVEC were incubated with unlabeled HK for varying times, washed, and the kinetics of BK release by plasma kallikrein were assayed by radioimmunoassay. Results indicated that kallikrein released BK from HUVEC in proportion to the initial amount of bound HK. No release of BK occurred in the absence of kallikrein. Also, there was no BK release upon kallikrein treatment of the HUVEC not treated with exogenous HK. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of HUVEC-bound 125I-HK indicated that addition of kallikrein resulted in cleavage of HK, thus corroborating the BK release experiments. Comparison of cleavage patterns has also indicated that cell-bound HK is slightly less susceptible to digestion by kallikrein than free HK. Therefore, our data suggest that human HK can bind to vascular endothelium in situ and that plasma kallikrein can recognize endothelial-bound HK as a substrate and liberate the vasoactive peptide BK.
- Published
- 1992
9. Intact Alzheimer amyloid precursor protein (APP) is present in platelet membranes and is encoded by platelet mRNA.
- Author
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Gardella JE, Ghiso J, Gorgone GA, Marratta D, Kaplan AP, Frangione B, and Gorevic PD
- Subjects
- Alzheimer Disease immunology, Amyloid beta-Peptides immunology, Amyloid beta-Protein Precursor, Antibodies immunology, Base Sequence, Blood Platelets drug effects, DNA chemistry, Endopeptidases pharmacology, Epitopes, Humans, Intracellular Membranes metabolism, Molecular Sequence Data, Polymerase Chain Reaction, Protein Precursors immunology, Thrombin pharmacology, Alzheimer Disease genetics, Amyloid beta-Peptides genetics, Blood Platelets chemistry, Protein Precursors genetics, RNA, Messenger biosynthesis
- Abstract
Using antibodies directed against N-terminal and C-terminal epitopes we have immunologically detected APP species in the membrane and saline-soluble fractions of unstimulated platelets, and in the conditioned medium of thrombin-stimulated platelets. These studies demonstrate an intact 140 kD membrane-associated form of APP that is released on degranulation. Evidence that platelets synthesize at least one form of APP (APP751) was obtained by enzymatic amplification of specific mRNA using Polymerase Chain Reaction (PCR) and direct sequence analysis of PCR product. Processing of APP for release may occur via successive C-terminal truncations, and/or by the release and proteolysis of an intact membrane associated form. An intact form of APP in platelets provides a circulating substrate upon which proteases from many tissues may act to produce beta protein (AB) during pathologic conditions.
- Published
- 1990
- Full Text
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10. Prevalence and functional role of anti-IgE autoantibodies in urticarial syndromes.
- Author
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Gruber BL, Baeza ML, Marchese MJ, Agnello V, and Kaplan AP
- Subjects
- Autoantibodies physiology, Basophils metabolism, Cold Temperature, Histamine Release, Humans, Immunoenzyme Techniques, Immunoglobulin G analysis, Immunoglobulin M analysis, Antibodies, Anti-Idiotypic analysis, Autoantibodies analysis, Immunoglobulin E immunology, Urticaria immunology
- Abstract
The prevalence of autoantibodies of immunoglobulin G (IgG) and immunoglobulin M (IgM) classes directed against myeloma immunoglobulin E (IgE) were determined in distinct subsets of urticaria, using an enzyme immunoassay. IgG anti-IgE antibodies were found in five of nine patients (55%) with cold urticaria, four of eight patients (50%) with urticarial vasculitis, and three of six patients (50%) with chronic urticaria. IgM anti-IgE antibodies were found exclusively in cold urticaria (two of nine patients, 22%). Heating of these sera increased the binding to IgE, suggesting immune complex formation. Several positive sera were capable of inducing histamine release from normal peripheral basophils and caused a wheal-flare response upon intradermal injection. Sera containing such autoantibodies from three cold urticaria patients were studied for passive transfer of cold sensitivity. One serum containing IgG anti-IgE gave a strongly positive transfer test at 5 h but not 48 h, suggesting a pathogenic role for the IgG.
- Published
- 1988
- Full Text
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11. The effect of C1 inhibitor upon Hageman factor autoactivation.
- Author
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Weiss R, Silverberg M, and Kaplan AP
- Subjects
- Electrophoresis, Polyacrylamide Gel, Factor XIIa, Humans, In Vitro Techniques, Kinetics, Temperature, Complement C1 Inactivator Proteins pharmacology, Factor XII analysis, Peptide Fragments analysis
- Abstract
Using components purified from human plasma, we have examined the effects of C1 inhibitor (C1 INH), the primary inhibitor of activated Hageman Factor (HFa) and Hageman factor fragment (HFf), on Hageman Factor (HF) autoactivation. When Hageman factor was exposed to a negatively charged surface, provided by either a glass cuvette or dextran sulfate, the addition of C1 INH gave a dose-dependent inhibition of the activity observed. The ability of C1 INH to decrease the maximal enzymatic activity generated was markedly temperature dependent with inhibition increasing as the temperature was raised from 4 degrees C to 37 degrees C. Although the rates of both autoactivation and inhibition were decreased at lower temperatures (4 degrees C), the latter rate was more sensitive to temperature modulation. When HF (final concentration 1 mumol/L) was incubated with C1 INH (0.54, 1.07, and 2.14 mumol/L) in the absence of an initiating surface, no increases in enzymatic activity were observed for up to 48 hours regardless of the C1 INH concentration. However, SDS polyacrylamide gel electrophoresis of the incubation mixture revealed that HF autodigestion had occurred by 48 hours despite the presence of C1 INH. In addition, the appearance of a new band suggested that a complex had been formed between the inhibitor and activated HF. Our findings indicate that C1 INH does not prevent HF autoactivation but rather inactivates the products of HF autodigestion.
- Published
- 1986
12. Contact activation of human plasma prorenin in vitro.
- Author
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Blumberg AL, Sealey JE, Atlas SA, Laragh JH, Dharmgrongartama B, and Kaplan AP
- Subjects
- Dialysis, Enzyme Activation, Factor XII Deficiency metabolism, Humans, Hydrogen-Ion Concentration, Kaolin pharmacology, Peptide Hydrolases metabolism, Prekallikrein metabolism, Enzyme Precursors blood, Renin blood
- Abstract
Acid activation of plasma prorenin occurs during dialysis to pH 3.3. and also during subsequent dialysis to pH 7.4. The latter, alkaline phase, involves Hageman factor-dependent formation of kallikrein, which in turn activates prorenin. The present study evaluates whether prorenin activation always occurs whenever kallikrein is activated in plasma. TAME esterase activity was used as a measure of plasma kallikrein activity an increase was observed during the alkaline phase of acid activation of prorenin. TAME esterase activity was absent when Hageman factor- or prekallikrein-deficient plasmas were similarly assayed and prorenin was not activated. Kaolin treatment of normal plasma rapidly increased TAME esterase activity at both 25 degrees and -4 degrees C, but no prorenin activation occurred. Similar changes in TAME esterase activity were observed in acid-treated plasma, in which setting prorenin was activated. No change in TAME esterase or renin activity occurred after addition of kaolin to acid-treated plasma deficient in Hageman factor; however, both enzymatic activities increased slightly in acidified prekallikrein-deficient plasma. Mixtures of these deficient plasmas exhibited normal kaolin activation of both TAME esterase and prorenin after acidification. Thus both Hageman factor and prekallikrein are needed for optimal contact activation of prorenin. These results demonstrate that prorenin activation does not always occur when active kallikrein is present in plasma. Prior acidification appears to be a prerequisite. Acidified prorenin may be more susceptible to cleavage; alternatively, competing substrates and/or inhibitors of kallikrein may be destroyed at acid pH, thereby permitting kallikrein to activate prorenin. Under normal conditions, activation of the plasma kallikrein-kinin system appears unlikely to result in activation of prorenin in vivo.
- Published
- 1981
13. The IgE-dependent release of a Hageman factor cleaving factor from human lung.
- Author
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Meier HL, Flowers B, Silverberg M, Kaplan AP, and Newball HH
- Subjects
- Antigens metabolism, Chromatography, Gel, Culture Techniques, Humans, Kallikreins metabolism, Kinetics, Lung immunology, Molecular Weight, Protease Inhibitors metabolism, Factor XII metabolism, Immunoglobulin E metabolism, Lung enzymology
- Abstract
Passively sensitized human lung fragments were shown to release a protease by an IgE-dependent mechanism which can cleave human Hageman factor (Coagulation Factor XII). This enzyme, lung Hageman factor cleaving factor, was partially purified by ion exchange chromatography and gel filtration and was shown to be a serine protease with an apparent molecular weight of 12,000-13,000. This protease appears to be unrelated to any known activator of Hageman factor by molecular weight and inhibition profile and was shown to be distinct from an IgE-dependent prekallikrein activator, as well as the kininogenase activity defined as basophil kallikrein of anaphylaxis. Although it appears marginally capable of activating Hageman factor, it rapidly cleaves and inactivates the activated form so that the net effect is a loss of activatable Hageman factor. The result suggests that diminished levels of Hageman factor that may be seen associated with IgE-dependent reactions can be due to digestion and depletion rather than activation, and other criteria for activation of the contact system must be employed.
- Published
- 1986
14. Assessment of Hageman factor activation in human plasma: quantification of activated Hageman factor-C1 inactivator complexes by an enzyme-linked differential antibody immunosorbent assay.
- Author
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Kaplan AP, Gruber B, and Harpel PC
- Subjects
- Complement C1 Inactivator Proteins immunology, Enzyme-Linked Immunosorbent Assay, Factor XII physiology, Factor XIIa, Humans, Isoantibodies, Kallikreins analysis, Kaolin, Partial Thromboplastin Time, Peptide Fragments physiology, Blood Coagulation, Complement C1 Inactivator Proteins physiology, Factor XII analysis, Peptide Fragments analysis
- Abstract
An enzyme-linked immunosorbent assay has been developed for the quantitation of activated Hageman factor-C1 inactivator (HF-C1 INH) complexes. Addition of increasing quantities of either of the major forms of activated Hageman factor (HFa or HFf) to normal plasma or to Hageman factor-deficient plasma leads to a dose-dependent increase in activated HF-C1 INH complexes. As little as 0.5 micrograms/mL of activated HF added to plasma can be detected, corresponding to activation of approximately 2% of plasma HF. The sensitivity of the assay is increased at least tenfold when complexes are formed in HF-deficient plasma, indicating competition between unactivated HF and activated HF-C1 INH complexes for binding to the antibody. Specificity is demonstrated in that addition of activated HF to hereditary angioedema plasma yields less than 1% of the activated HF-C1 INH complex formation obtained with normal plasma. Kaolin activation of HF-deficient plasma yields no detectable complex formation. Kaolin activation of prekallikrein-deficient plasma demonstrates a time-dependent increase in formation of activated HF-C1 INH complex consistent with the ability of HF in this plasma to autoactivate as the time of incubation with the surface is increased. Kaolin treatment of high-molecular weight (HMW) kininogen-deficient plasma yields an even more profound abnormality in the rate of formation of activated HF-C1 INH complexes reflecting the complex role of HMW kininogen in the initiation of contact activation. Although addition of corn inhibitor to plasma prevents activated HF-C1 INH complex formation, it does not inhibit activated HF sufficiently fast to prevent prekallikrein activation.
- Published
- 1985
15. Monoclonal antibody to human high-molecular-weight kininogen recognizes its prekallikrein binding site and inhibits its coagulant activity.
- Author
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Reddigari SR and Kaplan AP
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Monoclonal physiology, Antibody Affinity, Antibody Specificity, Binding, Competitive, Blood Coagulation Tests, Humans, Hydrolysis, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Peptide Fragments isolation & purification, Peptide Fragments pharmacology, Antibodies, Monoclonal immunology, Binding Sites, Antibody, Blood Coagulation, Kininogens immunology, Prekallikrein metabolism
- Abstract
We developed a mouse monoclonal antibody (MoAb 115-21) to human high-molecular-weight kininogen (HK) that recognizes its prekallikrein binding site (residues 565 through 595 of HK). The corresponding synthesized 31-amino acid peptide (peptide IV) was recently shown to retain native HK's prekallikrein binding property. The same peptide bound factor XI also, although less avidly. Our MoAb recognizes purified HK, peptide IV, and the light chain moiety of HK (where the peptide IV resides), as shown by enzyme-linked immunosorbent assay (ELISA) and Western blotting experiments. The apparent dissociation constant for the HK and MoAb 115-21 interaction was 2.2 nmol/L. It does not recognize low-molecular-weight kininogen (LK) with which HK shares its heavy chain moiety or any antigens in human plasma congenitally deficient in kininogens. The binding of MoAb 115-21 to purified light chain of HK was competitively inhibited by peptide IV. In addition, the antibody inhibits HK-dependent clotting activity of normal human plasma and dextran sulfate-mediated activation of prekallikrein in plasma and retards cleavage of HK in normal plasma after contact activation with dextran sulfate. Also, purified Fab fragments of MoAb 115-21 inhibited the HK-dependent coagulant activity and dextran sulfate-mediated prekallikrein activation in normal plasma. Since the kd for HK-MoAb 115-21 interaction is ten times lower than that of HK-prekallikrein, our data suggest that binding of MoAb 115-21 to HK's peptide IV site increases the free prekallikrein concentration in plasma and thus results in the decreased efficiency of factor XIIa-mediated activation of prekallikrein. Decreased levels of kallikrein thus formed may be responsible for the inhibition of HK-dependent clotting activity and the decrease in rate and extent of HK cleavage in normal plasma on contact activation with dextran sulfate. MoAb 115-21 may thus prove very useful, especially with its high affinity for HK, in further delineation of the role of HK and prekallikrein in contact activation and kinin-related human pathology.
- Published
- 1989
16. Hageman-factor-dependent fibrinolysis: generation of fibrinolytic activity by the interaction of human activated factor XI and plasminogen.
- Author
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Mandle RJ Jr and Kaplan AP
- Subjects
- Blood Coagulation Disorders enzymology, Factor XI Deficiency blood, Factor XII Deficiency blood, Humans, Plasminogen Activators immunology, Plasminogen Activators pharmacology, Prekallikrein, gamma-Globulins, Factor XI metabolism, Factor XII metabolism, Fibrinolysis, Plasminogen metabolism
- Abstract
Human coagulation factor XI has been purified, and upon activation with Hageman factor fragments, was found to convert the fibrinolytic proenzyme plasminogen to plasmin. This proactivator activity was shown to be functionally and antigenically distinct from prekallikrein. When the gamma-globulin fractions of plasma deficient in Hageman factor, prekallikrein and factor XI were isolated, factor-XI-deficient plasma possessed two-thirds of the plasminogen proactivator activity of the Hageman-factor-deficient plasma, while prekallikrein deficient plasma had only one-third of the plasminogen proactivator activity. Thus, the Hageman-factor-dependent plasminogen proactivator previously reported to be present in the gamma-globulin fraction of normal human plasma is a function of prekallikrein and factor XI, while the activity observed in prekallikrein-deficient plasma is attributable to factor XI. When compared utilizing digestion of iodinated fibrin, prekallikrein and factor XIa had similar potency per active site; they were, however, far less active than urokinase.
- Published
- 1979
17. Cleavage of human high-molecular weight kininogen by purified kallikreins and upon contact activation of plasma.
- Author
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Reddigari S and Kaplan AP
- Subjects
- Antibodies, Monoclonal immunology, Bradykinin biosynthesis, Dextran Sulfate, Dextrans, Humans, Kallikreins classification, Kaolin, Kininogens immunology, Molecular Weight, Kallikreins metabolism, Kininogens metabolism
- Abstract
To study the digestion pattern of human high-molecular weight (mol wt) kininogen (HMWK) in plasma during contact activation we have prepared monoclonal antibodies (MoAbs) to the light-chain (LC) and the heavy-chain moiety of HMWK. One MoAb from each set was purified, and neither MoAb inhibited the clotting activity of HMWK. In enzyme-linked immunosorbent assay and immunoblotting experiments neither antibody bound to kininogen-deficient plasma. Digestion of purified HMWK with plasma kallikrein yielded, on reduced sodium dodecyl sulfate gels, two LC forms, at 62 and 49 kd, respectively. Digestion of HMWK with tissue kallikrein (TK) yielded mainly the 62-kd form. In immunoblot analyses of these digests, the anti-LC MoAb detected products at 62 and 49 kd respectively. With plasma kallikrein, the 62-kd species slowly shifted to 49 kd, and with TK, the 62-kd species accumulated with time. Anti-LC MoAb was also used as a probe in immunoblotting experiments to study the digestion pattern of HMWK in whole plasma activated with kaolin or dextran sulfate. In activated normal pooled plasma (NHP) and factor XI-deficient plasma, native HMWK (mol wt, 115 kd) was cleaved within five to ten minutes, and two LC forms at 62 and 49 kd were detected. In kaolin-activated prekallikrein (PK)-deficient plasma, the disappearance of the 115-kd form was relatively slow, and only the 62-kd form of LC was seen. HMWK was not cleaved when factor XII-deficient plasma was incubated with kaolin. LC-dependent coagulant activity paralleled the presence of LC bands seen in the immunoblots, and lower-mol wt fragments of LC were not identified. These data indicate that in activated NHP two forms of LC of HMWK (62 and 49 kd) are formed sequentially. Further, the LC-dependent coagulant activity remains detectable long enough to suggest that proteolytic inactivation of LC is too slow to be an important control mechanism.
- Published
- 1988
18. Autoactivatability of human Hageman factor (factor XII).
- Author
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Miller G, Silverberg M, and Kaplan AP
- Subjects
- Biotransformation, Disulfides, Humans, Isoflurophate, Macromolecular Substances, Molecular Weight, Protein Binding, Factor XII metabolism
- Published
- 1980
- Full Text
- View/download PDF
19. In vivo studies of the pathogenesis of cold urticaria, cholinergic urticaria, and vibration-induced swelling.
- Author
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Kaplan AP and Beaven MA
- Subjects
- Angioedema etiology, Cold Temperature adverse effects, Histamine Release, Humans, Immunization, Passive, Parasympathetic Nervous System physiology, Urticaria immunology, Vibration adverse effects, Urticaria etiology
- Published
- 1976
- Full Text
- View/download PDF
20. Immunoreactive tissue kallikrein in human serum.
- Author
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Shimamoto K, Mayfield RK, Margolius HS, Chao J, Stroud W, and Kaplan AP
- Subjects
- Adult, Diet, Sodium-Restricted, Factor XII blood, Female, Humans, Kallikreins isolation & purification, Kallikreins urine, Male, Middle Aged, Pancreas analysis, Pancreatic Diseases blood, Prekallikrein blood, Saliva analysis, Statistics as Topic, Tissue Extracts analysis, Kallikreins blood, Radioimmunoassay
- Abstract
Human urinary kallikrein and an antiserum to it raised in the rabbit were used to detect and quantitate immunoreactive tissue kallikrein in human serum. Both 125I-labeled kallikrein and the unlabeled purified enzyme appear complexed to higher molecular weight entities in serum, but specific binding between radiolabeled enzyme and antiserum was unaffected by the presence of serum or plasma. Parallelism to standard displacement curves was always seen with radioimmunoassay of normal sera as well as with human mixed saliva or pancreatic extracts. Assay sensitivity is 160 pg/ml of serum, or 16 pg per tube. Purified plasma kallikrein or prekallikrein in concentrations up to 10 micrograms/ml showed no displacement. Acetone-kaolin activation of plasma produced the expected 30-fold increase in Tos-Arg-OMe esterase activity but no change in immunoreactive tissue kallikrein levels. Serum concentrations were 3.8 +/- 0.7 (mean +/- SE) ng/ml in 21 normal volunteers, and were similar in patients with Fletcher trait or Hageman factor deficiency. Significantly increased serum concentrations were seen with long-term low dietary sodium intake or acute forms of pancreatitis. Although the relation of this immunoreactive material to any active tissue kallikrein within the circulation remains to be determined, our studies provide a new parameter for the assessment of a system repeatedly suggested to have some role in regulation of vascular resistance.
- Published
- 1984
21. Quantification of human high molecular weight kininogen by immunoblotting with a monoclonal anti-light chain antibody.
- Author
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Reddigari S and Kaplan AP
- Subjects
- Animals, Blood Coagulation Tests, Humans, Kininogens immunology, Kininogens isolation & purification, Mice, Molecular Weight, Protein Conformation, Sensitivity and Specificity, Antibodies, Monoclonal, Immunoblotting methods, Kininogens blood
- Abstract
Activation of the Hageman factor-dependent pathways in human plasma leads to the cleavage of high molecular weight kininogen (HMWK) into a disulfide-linked two-chain (heavy and light chain) molecule and release of bradykinin, a vasoactive peptide. We have utilized murine monoclonal antibodies to the light chain of HMWK (Blood (1988) 71, 1344) and developed a very sensitive immunoblotting assay to detect and quantify the amount of cleaved or uncleaved HMWK in whole plasma. The total HMWK content of plasma from apparently healthy donors was 55 micrograms/ml by this method. Cleaved HMWK was detected when only 2% of the plasma had been activated and the method was sensitive down to 2 ng of HMWK. Because of the extreme lability of bradykinin in body fluids, quantification of cleaved HMWK provides an important adjunct which reflects contact activation and permits calculation of a theoretical upper limit of the potential kinin formed.
- Published
- 1989
- Full Text
- View/download PDF
22. The coagulation-kinin pathway of human plasma.
- Author
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Kaplan AP and Silverberg M
- Subjects
- Angioedema blood, Angioedema etiology, Blood Platelets physiology, Bradykinin metabolism, Factor XII physiology, Fibrinolysis, Humans, Inflammation blood, Inflammation etiology, Kininogens physiology, Prekallikrein physiology, Protein Precursors physiology, Protein Processing, Post-Translational, Blood Coagulation, Kinins physiology
- Published
- 1987
23. Enzymatic activities of activated and zymogen forms of human Hageman factor (factor XII).
- Author
-
Silverberg M and Kaplan AP
- Subjects
- Amides metabolism, Amino Acid Chloromethyl Ketones pharmacology, Electrophoresis, Polyacrylamide Gel, Enzyme Precursors pharmacology, Factor VIIa, Factor XIIa, Humans, Isoflurophate pharmacology, Kallikreins metabolism, Peptide Fragments, Plant Proteins pharmacology, Prekallikrein, Zea mays, Enzyme Precursors metabolism, Factor VII, Factor XII antagonists & inhibitors
- Abstract
Pro-Phe-Arg chloromethylketone (PPACMK) at 5.26 microM inactivated the amidolytic activity of native human Hageman factor with an apparent first-order rate constant of 0.75 min-1. The activated forms of Hageman factor, Hfa and HFf, were also inactivated by PPACMK with rate constants 0.82 and 0.72 min-1. These numbers indicate that the activity detectable in native Hageman factor is due to contamination with activated species. Uncleaved Hageman factor reacts slowly with 40 mM diisopropyl fluorophosphate with concomitant loss of its procoagulant activity. Incubation of native Hageman factor with PPACMK does not destroy its procoagulant activity, even in the presence of the activator dextran sulphate, but PPACMK inhibits autoactivation of Hageman factor, suggesting that no active site is formed in uncleaved, surface-bound Hageman factor. The activation of prekallikrein by Hageman factor under initial-rate conditions occurs after a lag and is prevented by an inhibitor of Hageman factor from corn. The kinetics of prekallikrein activation and the effects of inhibitors provide evidence that the amidolytic and proteolytic activities of human Hageman factor reside in the activated forms derived by limited proteolysis of the native molecule.
- Published
- 1982
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