DOCSLIB.ORG

Recombinant Human Plasma Kallikrein/KLKB1 Catalog Number: 2497-SE

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Recombinant Human Plasma Kallikrein/KLKB1 Catalog Number: 2497-SE Recombinant Human Plasma Kallikrein/KLKB1 Catalog Number: 2497-SE DESCRIPTION Source Mouse myeloma cell line, NS0-derived human Plasma Kallikrein/KLKB1 protein Gly20-Ala638, with a C-terminal 6-His tag Accession # P03952 N-terminal Sequence Gly20 Analysis Structure / Form Pro and processed forms Predicted Molecular 70 kDa Mass SPECIFICATIONS SDS-PAGE 80 kDa, reducing conditions Activity Measured by its ability to cleave a flourogenic peptide substrate Pro-Phe-Arg-7-amido-4-methylcoumarin (PFR-AMC). The specific activity is >1,000 pmol/min/µg, as measured under the described conditions. Endotoxin Level <1.0 EU per 1 μg of the protein by the LAL method. Purity >90%, by SDS-PAGE under reducing conditions and visualized by silver stain. Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl. See Certificate of Analysis for details. Activity Assay Protocol Materials Activation Buffer: 100 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 Assay Buffer: 50 mM Tris, 250 mM NaCl, pH 7.5 Recombinant Human Plasma Kallikrein/KLKB1 (rhKLKB1) (Catalog # 2497-SE) Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN) EDTA (Sigma, Catalog # E-4884) Substrate Pro-Phe-Arg-AMC (Bachem, Catalog # I-1295) F16 Black Maxisorp Plate (Nunc, Catalog # 475515) Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent Assay 1. Dilute rhKLKB1 to 200 µg/mL in Activation Buffer. 2. Dilute Thermolysin to 20 µg/mL in Activation Buffer. 3. Combine equal volumes of 200 µg/mL rhKLKB1 and 20 µg/mL Thermolysin for final concentrations of 100 µg/mL and 10 µg/mL respectively. 4. Incubate at 37 °C for 30 minutes. 5. Stop the reaction with 50 µM EDTA. 6. Dilute incubated rhKLKB1 to 0.5 ng/µL in Assay Buffer. 7. Dilute Substrate to 200 µM in Assay Buffer. 8. Load 50 μL of the 0.5 ng/µL rhKLKB1 in a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate. 9. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively in kinetic mode for 5 minutes. 10. Calculate specific activity: Adjusted Vmax * (RFU/min) x Conversion Factor** (pmol/RFU) Specific Activity (pmol/min/µg) = amount of enzyme (µg) *Adjusted for Substrate Blank **Derived using calibration standard, 7-amino-4-methylcoumarin (Sigma, Catalog # A9891). Final Assay Per Well: Conditions rhKLKB1: 0.025 µg Substrate: 100 µM PREPARATION AND STORAGE Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening. Rev. 5/20/2019 Page 1 of 2 Recombinant Human Plasma Kallikrein/KLKB1 Catalog Number: 2497-SE BACKGROUND Human Plasma Kallikrein, a serine protease, is synthesized in the liver and circulates in the plasma by binding to high molecular weight (HMW) kininogen or as a free zymogen. Once activated by its physiological activator, factor XII, it displays endopeptidase activity towards peptide bonds after arginine (preferred) and lysine. It cleaves HMW kininogen, its major physiological substrate, to release the potent vasodilator peptide bradykinin. It is also able to cleave a number of inactive precursor proteins to generate active products, such as plasminogen and prourokinase. Thus, it plays an important role in blood pressure regulation, fibrinolysis, and neutrophil activation (1). Human Plasma Kallikrein precursor contains a signal peptide (residues 1 to 19) and a pro form sequence (residues 20 to 638). Upon activation, the pro form is converted to a heavy chain and a light chain, which is linked by disulfide bonds and the latter contains the catalytic domain (2). The human Plasma Kallikrein pro form was expressed in the NS0 cells with a foreign signal peptide. The purified enzyme is mainly the pro form. When activated by thermolysin, it displays activity against a fluorogenic peptide substrate as described in Activity Assay Protocol. This activity can be inhibited by Human Serpin G1/C1 Inhibitor (Catalog # 2488-PI). References: 1. Coleman, R. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. (eds.) p. 1644, Academic Press, San Diego. 2. Chung, N.W. et al. (1986) Biochemistry 25:2410. Rev. 5/20/2019 Page 2 of 2 .
Load more

Recommended publications
  • Plasma Contact System Activation Drives Anaphylaxis in Severe Mast Cell–Mediated Allergic Reactions
    Plasma contact system activation drives anaphylaxis in severe mast cell–mediated allergic reactions Anna Sala-Cunill, MD, PhD,a,b,c Jenny Bjorkqvist,€ MSc,c,d Riccardo Senter, MD,c,e Mar Guilarte, MD, PhD,a,b Victoria Cardona, MD, PhD,a,b Moises Labrador, MD, PhD,a,b Katrin F. Nickel, PhD,c,d,f Lynn Butler, PhD,c,d,f Olga Luengo, MD, PhD,a,b Parvin Kumar, MSc,c,d Linda Labberton, MSc,c,d Andy Long, PhD,f Antonio Di Gennaro, PhD,c,d Ellinor Kenne, PhD,c,d Anne Jams€ a,€ PhD,c,d Thorsten Krieger, MD,f Hartmut Schluter,€ PhD,f Tobias Fuchs, PhD,c,d,f Stefanie Flohr, PhD,g Ulrich Hassiepen, PhD,g Frederic Cumin, PhD,g Keith McCrae, MD,h Coen Maas, PhD,i Evi Stavrou, MD,j and Thomas Renne, MD, PhDc,d,f Barcelona, Spain, Stockholm, Sweden, Padua, Italy, Hamburg, Germany, Basel, Switzerland, Cleveland, Ohio, and Utrecht, The Netherlands Background: Anaphylaxis is an acute, potentially lethal, hypotension. Activated mast cells systemically released heparin, multisystem syndrome resulting from the sudden release of mast which provided a negatively charged surface for factor XII cell–derived mediators into the circulation. autoactivation. Activated factor XII generates plasma Objectives and Methods: We report here that a plasma protease kallikrein, which proteolyzes kininogen, leading to the cascade, the factor XII–driven contact system, critically liberation of bradykinin. We evaluated the contact system in contributes to the pathogenesis of anaphylaxis in both murine patients with anaphylaxis. In all 10 plasma samples models and human subjects. immunoblotting revealed activation of factor XII, plasma Results: Deficiency in or pharmacologic inhibition of factor XII, kallikrein, and kininogen during the acute phase of anaphylaxis plasma kallikrein, high-molecular-weight kininogen, or the but not at basal conditions or in healthy control subjects.
    [Show full text]
  • SARS-Cov-2 Entry Protein TMPRSS2 and Its Homologue, TMPRSS4
    bioRxiv preprint doi: https://doi.org/10.1101/2021.04.26.441280; this version posted April 26, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 SARS-CoV-2 Entry Protein TMPRSS2 and Its 2 Homologue, TMPRSS4 Adopts Structural Fold Similar 3 to Blood Coagulation and Complement Pathway 4 Related Proteins ∗,a ∗∗,b b 5 Vijaykumar Yogesh Muley , Amit Singh , Karl Gruber , Alfredo ∗,a 6 Varela-Echavarría a 7 Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, México b 8 Institute of Molecular Biosciences, University of Graz, Graz, Austria 9 Abstract The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) utilizes TMPRSS2 receptor to enter target human cells and subsequently causes coron- avirus disease 19 (COVID-19). TMPRSS2 belongs to the type II serine proteases of subfamily TMPRSS, which is characterized by the presence of the serine- protease domain. TMPRSS4 is another TMPRSS member, which has a domain architecture similar to TMPRSS2. TMPRSS2 and TMPRSS4 have been shown to be involved in SARS-CoV-2 infection. However, their normal physiological roles have not been explored in detail. In this study, we analyzed the amino acid sequences and predicted 3D structures of TMPRSS2 and TMPRSS4 to under- stand their functional aspects at the protein domain level. Our results suggest that these proteins are likely to have common functions based on their conserved domain organization.
    [Show full text]
  • Activation of the Plasma Kallikrein-Kinin System in Respiratory Distress Syndrome
    003 I-3998/92/3204-043 l$03.00/0 PEDIATRIC RESEARCH Vol. 32. No. 4. 1992 Copyright O 1992 International Pediatric Research Foundation. Inc. Printed in U.S.A. Activation of the Plasma Kallikrein-Kinin System in Respiratory Distress Syndrome OLA D. SAUGSTAD, LAILA BUP, HARALD T. JOHANSEN, OLAV RPISE, AND ANSGAR 0. AASEN Department of Pediatrics and Pediatric Research [O.D.S.].Institute for Surgical Research. University of Oslo [L.B.. A.O.A.], Rikshospitalet, N-0027 Oslo 1, Department of Surgery [O.R.],Oslo City Hospital Ullev~il University Hospital, N-0407 Oslo 4. Department of Pharmacology [H. T.J.],Institute of Pharmacy, University of Oslo. N-0316 Oslo 3, Norway ABSTRAm. Components of the plasma kallikrein-kinin proteins that interact in a complicated way. When activated, the and fibrinolytic systems together with antithrombin 111 contact factors plasma prekallikrein, FXII, and factor XI are were measured the first days postpartum in 13 premature converted to serine proteases that are capable of activating the babies with severe respiratory distress syndrome (RDS). complement, fibrinolytic, coagulation, and kallikrein-kinin sys- Seven of the patients received a single dose of porcine tems (7-9). Inhibitors regulate and control the activation of the surfactant (Curosurf) as rescue treatment. Nine premature cascades. C1-inhibitor is the most important inhibitor of the babies without lung disease or any other complicating contact system (10). It exerts its regulatory role by inhibiting disease served as controls. There were no differences in activated FXII, FXII fragment, and plasma kallikrein (10). In prekallikrein values between surfactant treated and non- addition, az-macroglobulin and a,-protease inhibitor inhibit treated RDS babies during the first 4 d postpartum.
    [Show full text]
  • Human Plasma Kallikrein Releases Neutrophil Elastase During Blood Coagulation
    Human plasma kallikrein releases neutrophil elastase during blood coagulation. Y T Wachtfogel, … , A B Cohen, R W Colman J Clin Invest. 1983;72(5):1672-1677. https://doi.org/10.1172/JCI111126. Research Article Elastase is released from human neutrophils during the early events of blood coagulation. Human plasma kallikrein has been shown to stimulate neutrophil chemotaxis, aggregation, and oxygen consumption. Therefore, the ability of kallikrein to release neutrophil elastase was investigated. Neutrophils were isolated by dextran sedimentation, and elastase release was measured by both an enzyme-linked immunosorbent assay, and an enzymatic assay using t-butoxy-carbonyl-Ala- Ala-Pro-Val-amino methyl coumarin as the substrate. Kallikrein, 0.1-1.0 U/ml, (0.045-0.45 microM), was incubated with neutrophils that were preincubated with cytochalasin B (5 micrograms/ml). The release of elastase was found to be proportional to the kallikrein concentration. Kallikrein released a maximum of 34% of the total elastase content, as measured by solubilizing the neutrophils in the nonionic detergent Triton X-100. A series of experiments was carried out to determine if kallikrein was a major enzyme involved in neutrophil elastase release during blood coagulation. When 10 million neutrophils were incubated in 1 ml of normal plasma in the presence of 30 mM CaCl2 for 90 min, 2.75 micrograms of elastase was released. In contrast, neutrophils incubated in prekallikrein-deficient or Factor XII-deficient plasma released less than half of the elastase, as compared with normal plasma. The addition of purified prekallikrein to prekallikrein-deficient plasma restored neutrophil elastase release to normal levels.
    [Show full text]
  • Plasma Kallikrein Modulates Immune Cell Trafficking During Neuroinflammation Via PAR2 and Bradykinin Release
    Plasma kallikrein modulates immune cell trafficking during neuroinflammation via PAR2 and bradykinin release Kerstin Göbela,1,2, Chloi-Magdalini Asaridoua,2, Monika Merkera, Susann Eichlera, Alexander M. Herrmanna, Eva Geußb, Tobias Rucka, Lisa Schüngela,c, Linda Groenewega, Venu Narayanana, Tilman Schneider-Hohendorfa, Catharina C. Grossa, Heinz Wiendla, Beate E. Kehrelc, Christoph Kleinschnitzd,3, and Sven G. Meutha,3 aClinic of Neurology, Institute of Translational Neurology, University of Münster, 48149 Münster, Germany; bDepartment of Neurology, University Hospital Würzburg, 97080 Würzburg, Germany; cClinic of Anesthesiology, Intensive Care and Pain Medicine, Experimental and Clinical Haemostasis, University of Münster, 48149 Münster, Germany; and dDepartment of Neurology, University Hospital Essen, 45147 Essen, Germany Edited by Lawrence Steinman, Stanford University School of Medicine, Stanford, CA, and approved November 19, 2018 (received for review June 11, 2018) Blood–brain barrier (BBB) disruption and transendothelial trafficking peptidebradykinin(BK).Atthesametime,KKcanactivateFXIIin of immune cells into the central nervous system (CNS) are pathophys- a positive feedback loop, thereby leading to both activation of the iological hallmarks of neuroinflammatory disorders like multiple scle- intrinsic coagulation cascade and the proinflammatory KKS (14, rosis (MS). Recent evidence suggests that the kallikrein-kinin and 15). Additionally, KK may interact with cell-surface–associated re- coagulation system might participate in this process. Here, we iden- ceptors, such as the protease-activated receptors 1 (PAR1) and 2 tify plasma kallikrein (KK) as a specific direct modulator of BBB in- (PAR2) (16). tegrity. Levels of plasma prekallikrein (PK), the precursor of KK, were As KK has this dual mode of action (inflammation and co- markedly enhanced in active CNS lesions of MS patients.
    [Show full text]
  • Aeromonas Sobria a Serine Proteinase from Pressure Lowering Through Kinin Release by Induction of Vascular Leakage and Blood
    The Journal of Immunology Induction of Vascular Leakage and Blood Pressure Lowering through Kinin Release by a Serine Proteinase from Aeromonas sobria1 Takahisa Imamura,2* Hidemoto Kobayashi,‡ Rasel Khan,‡ Hidetoshi Nitta,*† and Keinosuke Okamoto‡ Aeromonas sobria causes septic shock, a condition associated with high mortality. To study the mechanism of septic shock by A. sobria infection, we examined the vascular leakage (VL) activity of A. sobria serine proteinase (ASP), a serine proteinase secreted by this pathogen. Proteolytically active ASP induced VL mainly in a bradykinin (BK) B2 receptor-, and partially in a histamine-H1 receptor-dependent manner in guinea pig skin. The ASP VL activity peaked at 10 min to 1.8-fold of the initial activity with an increased BK B2 receptor dependency, and attenuated almost completely within 30 min. ASP produced VL activity from human plasma apparently through kallikrein/kinin system activation, suggesting that ASP can generate kinin in humans. Consistent with the finding that a major part of the ASP-induced VL was reduced by a potent kallikrein inhibitor, soybean trypsin inhibitor that does not affect ASP enzymatic activity, ASP activated prekallikrein but not factor XII to generate kallikrein in a dose- and incubation time-dependent manner. ASP produced more VL activity directly from human low m.w. kininogen than high m.w. kininogen when both were used at their normal plasma concentrations. Intra- arterial injection of ASP into guinea pigs lowered blood pressure specifically via the BK B2 receptor. These data suggest that ASP induces VL through prekallikrein activation and direct kinin release from kininogens, which is a previously undescribed mechanism of A.
    [Show full text]
  • Assessing Plasmin Generation in Health and Disease
    International Journal of Molecular Sciences Review Assessing Plasmin Generation in Health and Disease Adam Miszta 1,* , Dana Huskens 1, Demy Donkervoort 1, Molly J. M. Roberts 1, Alisa S. Wolberg 2 and Bas de Laat 1 1 Synapse Research Institute, 6217 KD Maastricht, The Netherlands; [email protected] (D.H.); [email protected] (D.D.); [email protected] (M.J.M.R.); [email protected] (B.d.L.) 2 Department of Pathology and Laboratory Medicine and UNC Blood Research Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; [email protected] * Correspondence: [email protected]; Tel.: +31-(0)-433030693 Abstract: Fibrinolysis is an important process in hemostasis responsible for dissolving the clot during wound healing. Plasmin is a central enzyme in this process via its capacity to cleave fibrin. The ki- netics of plasmin generation (PG) and inhibition during fibrinolysis have been poorly understood until the recent development of assays to quantify these metrics. The assessment of plasmin kinetics allows for the identification of fibrinolytic dysfunction and better understanding of the relationships between abnormal fibrin dissolution and disease pathogenesis. Additionally, direct measurement of the inhibition of PG by antifibrinolytic medications, such as tranexamic acid, can be a useful tool to assess the risks and effectiveness of antifibrinolytic therapy in hemorrhagic diseases. This review provides an overview of available PG assays to directly measure the kinetics of plasmin formation and inhibition in human and mouse plasmas and focuses on their applications in defining the role of plasmin in diseases, including angioedema, hemophilia, rare bleeding disorders, COVID- 19, or diet-induced obesity.
    [Show full text]
  • The Role of the Plasminogen Activation System in Angioedema: Novel Insights on the Pathogenesis
    Journal of Clinical Medicine Review The Role of the Plasminogen Activation System in Angioedema: Novel Insights on the Pathogenesis Filomena Napolitano and Nunzia Montuori * Department of Translational Medical Sciences and Center for Basic and Clinical Immunology Research (CISI), University of Naples Federico II, 80135 Naples, Italy; fi[email protected] * Correspondence: [email protected]; Tel.: +39-0817463309 Abstract: The main physiological functions of plasmin, the active form of its proenzyme plasminogen, are blood clot fibrinolysis and restoration of normal blood flow. The plasminogen activation (PA) system includes urokinase-type plasminogen activator (uPA), tissue-type PA (tPA), and two types of plasminogen activator inhibitors (PAI-1 and PAI-2). In addition to the regulation of fibrinolysis, the PA system plays an important role in other biological processes, which include degradation of extracellular matrix such as embryogenesis, cell migration, tissue remodeling, wound healing, angiogenesis, inflammation, and immune response. Recently, the link between PA system and angioedema has been a subject of scientific debate. Angioedema is defined as localized and self- limiting edema of subcutaneous and submucosal tissues, mediated by bradykinin and mast cell mediators. Different forms of angioedema are linked to uncontrolled activation of coagulation and fibrinolysis systems. Moreover, plasmin itself can induce a potentiation of bradykinin production with consequent swelling episodes. The number of studies investigating the PA system involvement in angioedema has grown in recent years, highlighting its relevance in etiopathogenesis. In this review, we present the components and diverse functions of the PA system in physiology and its importance in angioedema pathogenesis. Citation: Napolitano, F.; Montuori, Keywords: angioedema; fibrinolysis; plasmin; uPAR; tPA N.
    [Show full text]
  • Human Tissue Kallikreins: a New Enzymatic Cascade Pathway?
    Biol. Chem., Vol. 383, pp. 1045 – 1057, July / August 2002 · Copyright © by Walter de Gruyter · Berlin · New York Review Human Tissue Kallikreins: A New Enzymatic Cascade Pathway? George M. Yousef and Eleftherios P. Diamandis* clude proteases which have an activated cysteine Department of Pathology and Laboratory Medicine, residue (cysteine proteases), aspartate (aspartate pro- Mount Sinai Hospital, Toronto, Ontario M5G 1X5, teases), metal ion (metalloproteases) or serine (serine Canada, and Department of Laboratory Medicine and proteases). Serine proteases are a family of enzymes that Pathobiology, University of Toronto, Toronto, Ontario, utilizes a uniquely activated serine residue in the sub- Canada strate-binding site to catalytically hydrolyze peptide bonds (Schultz and Liebman, 1997). This active site is characterized by the irreversible interaction with diiso- * Corresponding author propylfluorophosphate (DFP). Of all the serines in the protein, DFP can only react with the active serine to form Serine proteases are proteolytic enzymes with an ac- a phosphate ester (Schultz and Liebman, 1997). Out of tive serine residue in their catalytic site. Kallikreins are the estimated 400 – 500 proteases in the human genome, a subgroup of the serine protease family which is 32% are predicted to be serine proteases (Southan, known to have diverse physiological functions. The 2001). This large family includes the digestive enzymes human kallikrein gene family has now been fully char- (e.g., trypsin, chymotrtypsin), the kringle domain-con- acterized and includes 15 members tandemly located taining growth factors (e.g., tissue plasminogen activa- on chromosome 19q13.4. Here we discuss the com- tor), some of the blood clotting factors and the kallikreins.
    [Show full text]
  • Activation Profiles and Regulatory Cascades of the Human Kallikrein-Related Peptidases Hyesook Yoon
    Florida State University Libraries Electronic Theses, Treatises and Dissertations The Graduate School 2008 Activation Profiles and Regulatory Cascades of the Human Kallikrein-Related Peptidases Hyesook Yoon Follow this and additional works at the FSU Digital Library. For more information, please contact [email protected] FLORIDA STATE UNIVERSITY COLLEGE OF ARTS AND SCIENCES ACTIVATION PROFILES AND REGULATORY CASCADES OF THE HUMAN KALLIKREIN-RELATED PEPTIDASES By HYESOOK YOON A Dissertation submitted to the Department of Chemistry and Biochemistry in partial fulfillment of the requirements for the degree of Doctor of Philosophy Degree Awarded: Fall Semester, 2008 The members of the Committee approve the dissertation of Hyesook Yoon defended on July 10th, 2008. ________________________ Michael Blaber Professor Directing Dissertation ________________________ Hengli Tang Outside Committee Member ________________________ Brian Miller Committee Member ________________________ Oliver Steinbock Committee Member Approved: ____________________________________________________________ Joseph B. Schlenoff, Chair, Department of Chemistry and Biochemistry The Office of Graduate Studies has verified and approved the above named committee members. ii ACKNOWLEDGMENTS I would like to dedicate this dissertation to my parents for all your support, and my sister and brother. I would also like to give great thank my advisor, Dr. Blaber for his patience, guidance. Without him, I could never make this achievement. I would like to thank to all the members in Blaber lab. They are just like family to me and I deeply appreciate their kindness, consideration and supports. I specially like to thank to Mrs. Sachiko Blaber for her endless guidance and encouragement. I would like to thank Dr Jihun Lee, Margaret Seavy, Rani and Doris Terry for helpful discussions and supports.
    [Show full text]
  • Aprotinin Concentrations Effective for the Inihibition of Tissue Kallikrein
    KININS V PartB Edited by Keishi Abe Tohoku University School of Medicine Sendai, Miyagi, Japan Hiroshi Moriya Science University of Tokyo Tokyo, Japan and Setsuro Fujii The Osaka Foundation for Promotion of Fundamental Medical Research Otsu, Shiga, Japan PLENUM PRESS • NEW YORK AND LONDON CONTENTS - PART B KALLIKREIN AND OTHER PROTEASE INHIBITORS Human Kallistatin, a New Tissue Kallikrein-Binding Protein: Purification and Characterization 1 Maoyin Wang, Joseph Day, Lee Chao, and Julie Chao The Design of Specific Inhibitors of Tissue Kallikrein and Their Effect on the Blood Pressure of the Rat 9 James Burton and Athanassios Benetos Semisynthetic Arginine-15-Aprotinin, an Improved Inhibitor for Human Plasma Kallikrein 15 H. Tschesche, J. Beckmann, A. Mehlich, A. Feldmann, H.R. Wenzel, C.F. Scott, and R.W. Colman In Vivo Inhibition of Tissue Kallikreins by Kininogen Sequence Analogue Peptides 23 Hideki Okunishi, Jocelyn Spragg, James Burton, and Noboru Toda Highly Selective Synthetic Inhibitors with Regard to Plasma-Kallikrein Activities 29 S. Okamoto, U. Okamoto, K. Wanaka, A. Hikikata-Ikunomiya, M. Bohgaki, T. Naito, N. Horie, and Y. Okada Aprotinin Concentrations Effective for the Inhibition of Tissue Kallikrein and Plasma Kallikrein in Vitro and in Vivo 35 Hans Hoffmann, Matthias Siebeck, Olaf Thetter, Marianne Jochum, and Hans Fritz Changes in the Kallikrein-Kinin-System after Different Dose Regimen of Aprotinin During Cardiopulmonary Bypass Operation 43 W. Heller, G. Fuhrer, M.J. Gallimore, J. Michel, and H.-E. Hoffmeister Central Effect of Aprotinin, A Serine Protease Inhibitor, on Blood Pressure in Spontaneously Hypertensive and Wistar-Kyoto Rats 49 Shinji Seto, Masazumi Akahoshi, Shigeru Kusano, Shin-ichi Kitamura, and Kunitake Hashiba Augmentation of Kallikrein and Plasmin Inhibition Capacity by Aprotinin Using a New Assay to Monitor Therapy 55 M.J.
    [Show full text]
  • A Genomic Analysis of Rat Proteases and Protease Inhibitors
    A genomic analysis of rat proteases and protease inhibitors Xose S. Puente and Carlos López-Otín Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Instituto Universitario de Oncología, Universidad de Oviedo, 33006-Oviedo, Spain Send correspondence to: Carlos López-Otín Departamento de Bioquímica y Biología Molecular Facultad de Medicina, Universidad de Oviedo 33006 Oviedo-SPAIN Tel. 34-985-104201; Fax: 34-985-103564 E-mail: [email protected] Proteases perform fundamental roles in multiple biological processes and are associated with a growing number of pathological conditions that involve abnormal or deficient functions of these enzymes. The availability of the rat genome sequence has opened the possibility to perform a global analysis of the complete protease repertoire or degradome of this model organism. The rat degradome consists of at least 626 proteases and homologs, which are distributed into five catalytic classes: 24 aspartic, 160 cysteine, 192 metallo, 221 serine, and 29 threonine proteases. Overall, this distribution is similar to that of the mouse degradome, but significatively more complex than that corresponding to the human degradome composed of 561 proteases and homologs. This increased complexity of the rat protease complement mainly derives from the expansion of several gene families including placental cathepsins, testases, kallikreins and hematopoietic serine proteases, involved in reproductive or immunological functions. These protease families have also evolved differently in the rat and mouse genomes and may contribute to explain some functional differences between these two closely related species. Likewise, genomic analysis of rat protease inhibitors has shown some differences with the mouse protease inhibitor complement and the marked expansion of families of cysteine and serine protease inhibitors in rat and mouse with respect to human.
    [Show full text]