DOCSLIB.ORG

A Cysteine Proteinase Inhibitor of Human Saliva We Have Recently

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Cysteine Proteinase Inhibitor of Human Saliva We Have Recently COMMUNICATION J. Biochem. 96,1311-1314 (1984) Cystatin S : A Cysteine Proteinase Inhibitor of Human Saliva Satoko ISEMURA,* Eiichi SAITOH,* Seiki ITO,** Mamoru ISEMURA,*** and Kazuo SANADA* *Department of Oral Biochemistry , Niigata Faculty, Nippon Dental University. Niigata 951, **First Department of Internal Medicine, Niigata University School of Medicine, Niigata 951, and ***Department of Biochemistry, Tohoku University School of Medicine, Sendai, Miyagi 980 Received for publication, July 12, 1984 An acidic protein of human saliva, which we named SAP-1 previously, is now shown to be an inhibitor of several cysteine proteinases. The protein inhibited papain and ficin strongly, and stem bromelain and bovine cathepsin C partially. How ever, it did not inhibit either porcine cathepsin B or clostripain. The mode of the inhibition of papain was found to be non-competitive. The name cystatin S has been proposed for this salivary protein in view of the similarities in activity and structure to other cysteine proteinase inhibitors such as chicken egg-white cystatin and human cystatins A, B, and C. The cystatin S antigen was detected immunohistochemically in the serous cells of human parotid and submaxillary glands. We have recently isolated an acidic protein, SAP-1, 3.4.22.3], bromelain [EC 3.4.22.4], and clostripain with a molecular weight of 12,552 and PI 4.68 [EC 3.4.22.8] from Sigma; cathepsin C [dipeptidyl from human whole saliva, and determined its peptidase I, EC 3.4.14.1] from Serva; trypsin [EC amino acid sequence (1). This protein has se 3.4.21.4] and chymotrypsin [EC 3.4.21.1] from quence homology of 54% with human y-trace, the Worthington Biochemical Inc. Cathepsin B [EC cysteine proteinase inhibitor first found in cere 3.4.22.1] was prepared from porcine liver accord brospinal fluid (2, 3), and of 41 % with cystatin, ing to the method described by Takahashi et al. the cysteine proteinase inhibitor from egg-white (6). (4, 5). The high degree of structural similarity The activities of papain, ficin, bromelain, and has suggested that SAP-1 may be active in inhibit cathepsin B were measured with N-benzoyl-DL ing certain cysteine proteinases. In the present - arginine p-nitroanilide as a substrate according to work, we have found that is the case. We also the method described by Barrett (7). Cathepsin demonstrate that the serous cells of human parotid C was assayed by quantifying glycine released from and submaxillary glands are positively stained by the substrate Gly-Gly-Gly (Protein Research the indirect immunofluorescence method using the Foundation, Osaka). Trypsin and chymotrypsin antiserum raised against SAP-1. were assayed by the method of Schwert and Take Whole saliva was collected and SAP-1 was naka (8), and clostripain by the method of Mitchell isolated as described previously (1). The follow and Harrington (9). One unit (U) of enzyme was ing enzymes were obtained from the commercial defined as the activity hydrolyzing 1 ƒÊmol of the sources indicated: papain [EC 3.4.22.2], ficin [EC substrate per min under the conditions used. Vol. 96, No. 4, 1984 1311 1312 COMMUNICATION For the inhibition assay, papain, ficin, bro melain, clostripain, trypsin, and chymotrypsin were separately preincubated with varying amounts of SAP-1 in 500 ƒÊd of 0.05 M Tris-HCl buffer (pH 8.0) at 25•Ž for 5 min. In the cases of cathepsins B and C, preincubation with SAP-1 was performed in 500 ƒÊl of 0.05 M phosphate buffer (pH 6.5) at 25•Ž for 5 min and in 200,ƒÊl of 0.15 M acetate buffer (pH 4.7) containing 2.5 mm cysteine and 10 mm NaCl at 40•Ž for 5 min, respectively. The enzyme activity was then measured according to the method described above. In the case of ca Fig. 1. Inhibitory effect of SAP-1 on cysteine pro thepsin C 5 µl of 0.1 M Gly-Gly-Gly was added to teinases. Enzymes were preincubated for 5 min with the preincubated mixture of the enzyme and SAP varying amounts of the inhibitor and then the enzyme 1, and then the whole mixture was incubated for activities were determined as described in the text. 30 min at 40•Ž. The reaction was stopped by the Total reaction volumes were 2,050ƒÊl for assay of ca thepsin B, papain ficin and bromelain, or 205 ƒÊl for addition of 0.02% of monoiodoacetic acid in a that of cathepsin C. The enzymes used were 14.2 mU sampling buffer (2 ml) used for the amino acid of papain (•œ), 8.7 mU of ficin (•¢), 5.6 mU of bro analysis. Released glycine was quantified with a melain (•¡), 12.9 mU of cathepsin B (•›), and 1.7 mU Waters amino acid analysis system. of cathepsin C (•£). Results are expressed as % of SAP-1 did not show any activity inhibiting the residual activity of I mU of each enzyme and the serine proteinases, trypsin (1.69 U, 100 jig), and plotted against the amounts of the inhibitor. No in chymotrypsin (0.22 U, 100 jig) up to the amount hibition was observed when 0.7 U of clostripain, 1.69 U of 222 ,ƒÊg of SAP-1. The cysteine proteinases, of trypsin, or 0.22 U of chymotrypsin was tested. cathepsin B (12.9 mU) and clostripain (0.7 U 100 ƒÊ g) were not affected by SAP-1 (222 ƒÊg), either The cysteine proteinase inhibitors display con (Fig. 1). However, papain and ficin were inhib siderable variety in their relative activities from ited strongly, and bromelain and cathepsin C enzyme to enzyme on which they act (13, 16). partially (Fig. 1). The amounts of SAP-1 required As shown above, cystatin S did not inhibit cathep for 50% inhibition of the enzyme activity of 1 mU sin B. Thus cystatin S may be distinguished from of papain (2.18 jig), ficin (6.61 ƒÊg), and bromelain egg-white cystatin and human cystatins A, B, and (117.9 ƒÊg) were 0.8, 1.2, and 9.5 ƒÊg, respectively. C which inhibit human cathepsin B, although the When SAP-1 was reduced and S-carboxymeth possibility cannot be excluded that the difference ylated as described previously (1), the papain in may be due to the use of cathepsin B of different hibitory activity was lost completely. Thus the species. Therefore, the variety of inhibitions by conformation of the protein maintained by the inhibitors constituting a cystatin family appears disulfide bridges must be important for the ac to be much wider than has been considered (3). tivity. When the initial velocity of the reaction After the present work was completed, a paper with papain under various substrate concentrations appeared describing the presence of multiple forms in the presence or absence of SAP-1 was studied, of cysteine proteinase inhibitors in human saliva a Lineweaver-Burk plot demonstrated that SAP-I and salivary glands (17). Therefore, cystatin S is inhibited the enzyme non-competitively. considered to represent one of the several salivary These findings clearly indicate that SAP-1 is cysteine proteinase inhibitors, and the first with a a cysteine proteinase inhibitor. Several kinds of known amino acid sequence among them. cysteine proteinase inhibitor are now known (2 In the following experiments, sections of vari 5, 10-16) and some of them have been grouped ous human tissues were examined for the presence into a cystatin family (3). Because of the simi of cystatin S antigen. The rabbit antiserum larities in the amino acid sequence (1) and activity against cystatin S was prepared by the conven as revealed here between SAP-1 and cystatins, we tional method (16). The antiserum thus prepared propose the name cystatin S for SAP-l. formed a single precipitin line with cystatin S and J. Biochem. CYSTATIN S 1313 Fig. 2. Immunohistochemical detection of the cystatin S antigen in human tissues . Serial sections of parotid glands (A) or submaxillary glands (B) were stained with the anti-cystatin S antiserum (a), with the antiserum which had been absorbed with cystatin S (b), or with hematoxylin-eosin (c) (•~45 for (A) and •~90 for (B)) . The serous cells were positively stained. whole saliva, but not with human serum in an was not detected in these sections. The results Ouchterlony double immunodiffusion system. of the immunohistochemical examination may ex Histochemical detection by an indirect immu clude the possibility that cystatin S is the product nofluorescence technique and by hematoxylin of bacteria or exfoliated epithelial cells which are eosin staining were performed according to the also present in whole saliva. method described previously (18). Figure 2 shows Although the biological role of cystatin S is immunohistochemical evidence for the presence of not clear at present, these findings suggest that the cystatin S antigen in the serous cells of pa salivary glands, ducts or saliva may contain en rotid and submaxillary glands, suggesting that these zymes which are under regulation by the cysteine cells are responsible for its production. Sections proteinase inhibitor. It is also possible that the of pancreas and bronchea were also examined, inhibitor may function as protection for oral tis because these tissues have been shown to contain sues and proteins by inhibiting the enzymes pro the other salivary component, proline-rich peptide duced by oral bacteria. P-C (18, 19). However , the cystatin S antigen Vol. 96, No. 4, 1984 1314 COMMUNICATION 11. Takio, K., Kominami, E., Wakamatsu, N., Katsu numa, N., and Titani, K. (1983) Biochem. Biophys. REFERENCES Res. Common. 115, 902-908 1. Isemura, S., Saitoh, E., & Sanada, K. (1984) J. 12.
Load more

Recommended publications
  • Enzymes Handling/Processing
    Enzymes Handling/Processing 1 Identification of Petitioned Substance 2 3 This Technical Report addresses enzymes used in used in food processing (handling), which are 4 traditionally derived from various biological sources that include microorganisms (i.e., fungi and 5 bacteria), plants, and animals. Approximately 19 enzyme types are used in organic food processing, from 6 at least 72 different sources (e.g., strains of bacteria) (ETA, 2004). In this Technical Report, information is 7 provided about animal, microbial, and plant-derived enzymes generally, and more detailed information 8 is presented for at least one model enzyme in each group. 9 10 Enzymes Derived from Animal Sources: 11 Commonly used animal-derived enzymes include animal lipase, bovine liver catalase, egg white 12 lysozyme, pancreatin, pepsin, rennet, and trypsin. The model enzyme is rennet. Additional details are 13 also provided for egg white lysozyme. 14 15 Chemical Name: Trade Name: 16 Rennet (animal-derived) Rennet 17 18 Other Names: CAS Number: 19 Bovine rennet 9001-98-3 20 Rennin 25 21 Chymosin 26 Other Codes: 22 Prorennin 27 Enzyme Commission number: 3.4.23.4 23 Rennase 28 24 29 30 31 Chemical Name: CAS Number: 32 Peptidoglycan N-acetylmuramoylhydrolase 9001-63-2 33 34 Other Name: Other Codes: 35 Muramidase Enzyme Commission number: 3.2.1.17 36 37 Trade Name: 38 Egg white lysozyme 39 40 Enzymes Derived from Plant Sources: 41 Commonly used plant-derived enzymes include bromelain, papain, chinitase, plant-derived phytases, and 42 ficin. The model enzyme is bromelain.
    [Show full text]
  • Biochemistry and the Genomic Revolution 1.1
    Dedication About the authors Preface Tools and Techniques Clinical Applications Molecular Evolution Supplements Supporting Biochemistry, Fifth Edition Acknowledgments I. The Molecular Design of Life 1. Prelude: Biochemistry and the Genomic Revolution 1.1. DNA Illustrates the Relation between Form and Function 1.2. Biochemical Unity Underlies Biological Diversity 1.3. Chemical Bonds in Biochemistry 1.4. Biochemistry and Human Biology Appendix: Depicting Molecular Structures 2. Biochemical Evolution 2.1. Key Organic Molecules Are Used by Living Systems 2.2. Evolution Requires Reproduction, Variation, and Selective Pressure 2.3. Energy Transformations Are Necessary to Sustain Living Systems 2.4. Cells Can Respond to Changes in Their Environments Summary Problems Selected Readings 3. Protein Structure and Function 3.1. Proteins Are Built from a Repertoire of 20 Amino Acids 3.2. Primary Structure: Amino Acids Are Linked by Peptide Bonds to Form Polypeptide Chains 3.3. Secondary Structure: Polypeptide Chains Can Fold Into Regular Structures Such as the Alpha Helix, the Beta Sheet, and Turns and Loops 3.4. Tertiary Structure: Water-Soluble Proteins Fold Into Compact Structures with Nonpolar Cores 3.5. Quaternary Structure: Polypeptide Chains Can Assemble Into Multisubunit Structures 3.6. The Amino Acid Sequence of a Protein Determines Its Three-Dimensional Structure Summary Appendix: Acid-Base Concepts Problems Selected Readings 4. Exploring Proteins 4.1. The Purification of Proteins Is an Essential First Step in Understanding Their Function 4.2. Amino Acid Sequences Can Be Determined by Automated Edman Degradation 4.3. Immunology Provides Important Techniques with Which to Investigate Proteins 4.4. Peptides Can Be Synthesized by Automated Solid-Phase Methods 4.5.
    [Show full text]
  • Evidence for an Active-Center Cysteine in the SH-Proteinase Cu-Clostripain Through Use of IV-Tosyl-L-Lysine Chloromethyl Ketone
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Volume 173, number 1 FEBS 1649 July 1984 Evidence for an active-center cysteine in the SH-proteinase cu-clostripain through use of IV-tosyl-L-lysine chloromethyl ketone A.-M. Gilles and B. Keil Unitt! de Chimie des Protknes, Institut Pasteur, 28, rue du Docteur Roux, 75724 Paris CPdex 15, France Received 30 May 1984 The rapid reaction of a-clostripain with tosyl-L-lysine chloromethyl ketone results in a complete loss of activity and in the disappearance of one titratable SH group whereas the number of histidine residues is not affected. Tosyl-L-phenylalanine chloromethyl ketone and phenylmethylsulfonyl fluoride have no effect on the catalytic activity. From the molar ratio and under the assumption of 1: 1 molar interaction, the fully active enzyme has a specific activity of 650-700 units/mg [twice the value proposed by Porter et al. (J. Biol. Chem. 246 (1971) 76757682)]. Partial oxidation makes it experimentally impossible to attain this maximal value. ff-Clostripain Cysteine proteinase Active site 1. INTRODUCTION was due to the modification of a thiol group in an analogous way with other cysteine proteinases such Clostripain (EC 3.4.4.20) is a sulfhydryl protein- as papain [6] and ficin [7]. Recently [8], we eluci- ase isolated from the culture filtrate of Clostridium dated the amino acid sequence around this acces- histolyticum with a highly limited specificity sible thiol group after labelling with radioactive directed at the carboxyl bond of arginyl residues in iodoacetic acid.
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • The Proteolysis of Apolipoprotein E in Alzheimer's Disease
    THE PROTEOLYSIS OF APOLIPOPROTEIN E IN ALZHEIMER’S DISEASE by Julia Love A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Biology Boise State University August 2016 © 2016 Julia Love ALL RIGHTS RESERVED BOISE STATE UNIVERSITY GRADUATE COLLEGE DEFENSE COMMITTEE AND FINAL READING APPROVALS of the thesis submitted by Julia Love Thesis Title: The Proteolysis of Apolipoprotein E in Alzheimer’s Disease Date of Final Oral Examination: 26 April 2016 The following individuals read and discussed the thesis submitted by student Julia Love, and they evaluated her presentation and response to questions during the final oral examination. They found that the student passed the final oral examination. Troy Rohn, Ph.D. Chair, Supervisory Committee Kenneth A. Cornell, Ph.D. Member, Supervisory Committee Juliette Tinker, Ph.D. Member, Supervisory Committee The final reading approval of the thesis was granted by Troy Rohn, Ph.D., Chair of the Supervisory Committee. The thesis was approved for the Graduate College by Jodi Chilson, M.F.A., Coordinator of Theses and Dissertations. DEDICATION This thesis is dedicated to my parents Paul and Cynthia Love, my brother Philip Love, and all of my friends who have supported and encouraged me along the way. iv ACKNOWLEDGEMENTS There have been many people who have contributed to this work and my academic growth over the course of pursuing my Master’s degree. These individual contributions have not gone unnoticed and are an important part of my thesis work. First and foremost, I would like to thank Dr. Troy Rohn for being available with a willing attitude whenever I needed assistance, for his steadfast support and care, and for providing me with every opportunity to exceed what I thought were my limitations.
    [Show full text]
  • Molecular Insights Into Bromelain Application in Industry and Health Care
    Biosc.Biotech.Res.Comm. Special Issue Vol 13 No 15 (2020) Pp-36-46 Molecular Insights into Bromelain Application in Industry and Health Care Sushma S. Murthy and T. Bala Narsaiah 1Research Scholar, Department of Chemical Engineering, JNTUA College of Engineering, Ananthapuram-515002, Andhra Pradesh, India 2Department of Chemical Engineering, JNTUA College of Engineering, Ananthapuram-515002, Andhra Pradesh, India ABSTRACT Bromelain is a cysteine protease derived from the stem and fruit of the pineapple. It has a significant role in pharmacological and clinical applications. Studies have shown Bromelain to be a potent photoactive compound that has a wide application in industry. It has also been shown to be effective in treatment of cancer, inflammation, and allergies. It has a distinct immunomodulatory activity which forms an important strategy in its utilization as a therapeutic agent. Bromelain plays a significant role at molecular level by regulating the expression of proteins that are potential therapeutic targets. Bromelain is used extensively worldwide as an herbal medicine as it promises good efficacy and has no side effects. This paper reviews the general characteristics of Bromelain, its separation process, and its use in industries and healthcare as a therapeutic agent. The present review identifies that there is lack of knowledge pertaining to the mode of action of Bromelain in inhibiting transcriptional factors and in controlling cancer. An in-detail analysis in this area might help in expanding the therapeutic scope of Bromelain. KEY WORDS: BROMELAIN, CANCER, PHARMacOLOGICAL actiVITY, SEpaRatiON, TRANSCRIPTION FactORS. INTRODUCTION because of its wide benefits to the human system. The stem part of the plant, which is inexpensive and is usually The Bromelain protease is isolated from the stem and discarded, has a high concentration of bromelain and fruit of Pineapple (Ananas comosus,).
    [Show full text]
  • Proteolytic Enzymes in Grass Pollen and Their Relationship to Allergenic Proteins
    Proteolytic Enzymes in Grass Pollen and their Relationship to Allergenic Proteins By Rohit G. Saldanha A thesis submitted in fulfilment of the requirements for the degree of Masters by Research Faculty of Medicine The University of New South Wales March 2005 TABLE OF CONTENTS TABLE OF CONTENTS 1 LIST OF FIGURES 6 LIST OF TABLES 8 LIST OF TABLES 8 ABBREVIATIONS 8 ACKNOWLEDGEMENTS 11 PUBLISHED WORK FROM THIS THESIS 12 ABSTRACT 13 1. ASTHMA AND SENSITISATION IN ALLERGIC DISEASES 14 1.1 Defining Asthma and its Clinical Presentation 14 1.2 Inflammatory Responses in Asthma 15 1.2.1 The Early Phase Response 15 1.2.2 The Late Phase Reaction 16 1.3 Effects of Airway Inflammation 16 1.3.1 Respiratory Epithelium 16 1.3.2 Airway Remodelling 17 1.4 Classification of Asthma 18 1.4.1 Extrinsic Asthma 19 1.4.2 Intrinsic Asthma 19 1.5 Prevalence of Asthma 20 1.6 Immunological Sensitisation 22 1.7 Antigen Presentation and development of T cell Responses. 22 1.8 Factors Influencing T cell Activation Responses 25 1.8.1 Co-Stimulatory Interactions 25 1.8.2 Cognate Cellular Interactions 26 1.8.3 Soluble Pro-inflammatory Factors 26 1.9 Intracellular Signalling Mechanisms Regulating T cell Differentiation 30 2 POLLEN ALLERGENS AND THEIR RELATIONSHIP TO PROTEOLYTIC ENZYMES 33 1 2.1 The Role of Pollen Allergens in Asthma 33 2.2 Environmental Factors influencing Pollen Exposure 33 2.3 Classification of Pollen Sources 35 2.3.1 Taxonomy of Pollen Sources 35 2.3.2 Cross-Reactivity between different Pollen Allergens 40 2.4 Classification of Pollen Allergens 41 2.4.1
    [Show full text]
  • Durham E-Theses
    Durham E-Theses Midgut proteases from larval spodoptera littoralis (lepidoptera: noctutoae) Lee, Michael James How to cite: Lee, Michael James (1992) Midgut proteases from larval spodoptera littoralis (lepidoptera: noctutoae), Durham theses, Durham University. Available at Durham E-Theses Online: http://etheses.dur.ac.uk/5739/ Use policy The full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-prot purposes provided that: • a full bibliographic reference is made to the original source • a link is made to the metadata record in Durham E-Theses • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders. Please consult the full Durham E-Theses policy for further details. Academic Support Oce, Durham University, University Oce, Old Elvet, Durham DH1 3HP e-mail: [email protected] Tel: +44 0191 334 6107 http://etheses.dur.ac.uk MIDGUT PROTEASES FROM LARVAL SPODOPTERA LITTORALIS (LEPIDOPTERA: NOCTUTOAE) By Michael James Lee B.Sc. (Dunelm) The copyright of this thesis rests with the author. No quotation from it should be pubhshed without his prior written consent and information derived from it should be acknowledged. Being a thesis submitted for the degree of Doctor of Philosophy of the University of Durham. November, 1992 Hatfield College University of Durham 6 APR 1993 DECLARATION I hereby declare that the work presented in this document is based on research carried out by me, and that no part has been previously submitted for a degree in this or any other university.
    [Show full text]
  • Role of Systemic Enzymes in Infections
    Article ID: WMC002495 2046-1690 Role of Systemic Enzymes in Infections Corresponding Author: Dr. Sukhbir Shahid, Consultant Pediatrician, Pediatrics - India Submitting Author: Dr. Sukhbir Shahid, Consultant Pediatrician, Pediatrics - India Article ID: WMC002495 Article Type: Review articles Submitted on:22-Nov-2011, 08:16:56 AM GMT Published on: 22-Nov-2011, 02:34:00 PM GMT Article URL: http://www.webmedcentral.com/article_view/2495 Subject Categories:COMPLEMENTARY MEDICINE Keywords:Enzymes, Systemic enzymes, Infections, Sepsis, Proteolytic, Supplementary How to cite the article:Shahid S . Role of Systemic Enzymes in Infections . WebmedCentral COMPLEMENTARY MEDICINE 2011;2(11):WMC002495 Source(s) of Funding: None Competing Interests: None WebmedCentral > Review articles Page 1 of 13 WMC002495 Downloaded from http://www.webmedcentral.com on 23-Dec-2011, 07:57:46 AM Role of Systemic Enzymes in Infections Author(s): Shahid S Abstract infections[4]. The ‘battle’ between the host’s immunity and organism leads to a lot of ‘molecular’morbidity and mortality. Anti-infective agents do help but at times benefit is marginal. These agents may sometimes Enzymes are complex macromolecules of amino-acids worsen the situation through release of immune which bio-catalyse various body processes. Adequate complexes and dead bacilli into the blood stream. concentrations of enzymes are essential for optimal They also fail to reverse the hemodynamic instability functioning of the immune system. During infections, and immune paralysis characteristic of these body’s enzymatic system is attacked and hence the infections[4]. Supplementation with drugs targeted immune system is also likely to derange. This may be against this ‘choatic’ or ‘dysfunctional’ immune detrimental for the host’s well-being and existence.
    [Show full text]
  • Concentrate of Proteolytic Enzymes Enriched in Bromelain
    20 September 2012 EMA/648483/2012 Committee for Medicinal Products for Human Use (CHMP) Assessment report NexoBrid Concentrate of proteolytic enzymes enriched in bromelain Procedure No. EMEA/H/C/002246 Note Assessment report as adopted by the CHMP with all information of a commercially confidential nature deleted. 7 Westferry Circus ● Canary Wharf ● London E14 4HB ● United Kingdom Telephone +44 (0)20 7418 8400 Facsimile +44 (0)20 7418 8416 E-mail [email protected] Website www.ema.europa.eu An agency of the European Union © European Medicines Agency, 2012. Reproduction is authorised provided the source is acknowledged. Table of contents 1. Background information on the procedure .............................................. 5 1.1. Submission of the dossier.................................................................................... 5 1.2. Steps taken for the assessment of the product ....................................................... 6 2. Scientific discussion ................................................................................ 7 2.1. Introduction ...................................................................................................... 7 2.2. Quality aspects .................................................................................................. 9 2.3. Non-clinical aspects .......................................................................................... 20 2.4. Clinical aspects ................................................................................................ 29 2.5.
    [Show full text]
  • Enzymes for Cell Dissociation and Lysis
    Issue 2, 2006 FOR LIFE SCIENCE RESEARCH DETACHMENT OF CULTURED CELLS LYSIS AND PROTOPLAST PREPARATION OF: Yeast Bacteria Plant Cells PERMEABILIZATION OF MAMMALIAN CELLS MITOCHONDRIA ISOLATION Schematic representation of plant and bacterial cell wall structure. Foreground: Plant cell wall structure Background: Bacterial cell wall structure Enzymes for Cell Dissociation and Lysis sigma-aldrich.com The Sigma Aldrich Web site offers several new tools to help fuel your metabolomics and nutrition research FOR LIFE SCIENCE RESEARCH Issue 2, 2006 Sigma-Aldrich Corporation 3050 Spruce Avenue St. Louis, MO 63103 Table of Contents The new Metabolomics Resource Center at: Enzymes for Cell Dissociation and Lysis sigma-aldrich.com/metpath Sigma-Aldrich is proud of our continuing alliance with the Enzymes for Cell Detachment International Union of Biochemistry and Molecular Biology. Together and Tissue Dissociation Collagenase ..........................................................1 we produce, animate and publish the Nicholson Metabolic Pathway Hyaluronidase ...................................................... 7 Charts, created and continually updated by Dr. Donald Nicholson. DNase ................................................................. 8 These classic resources can be downloaded from the Sigma-Aldrich Elastase ............................................................... 9 Web site as PDF or GIF files at no charge. This site also features our Papain ................................................................10 Protease Type XIV
    [Show full text]
  • Degradative Inactivation of Cyclic AMP-Dependent Protein
    Proc. Natl Acad. Sci. USA Vol. 78, No. 6, pp. 3492-3495, June 1981 Biochemistry Degradative inactivation of cyclic AMP-dependent protein kinase by a membranal proteinase is restricted to the free catalytic subunit in its native conformation (brush-border membranes/intestinal microvilli/enzyme regulation/limited proteolysis) EYTAN ALHANATY, JONATHAN PATINKIN, MIRIAM TAUBER-FINKELSTEIN, AND SHMUEL SHALTIEL* Department of Chemical Immunology, The Weizmann Institute of Science, Rehovot, Israel Communicated by Michael Sela, March 13, 1981 ABSTRACT A membranal proteinase from brush-border ep- This raises the possibility that there may be additional regula- ithelial cells of the rat small intestine was shown to bring about a tory devices for modulating the cellular response to the hor- restricted and limited degradation ofthe free catalytic subunit (C) monal stimulus. of cyclic AMP-dependent protein kinase (ATP:protein phospho- cAMPdPKase activity in brush-border membranes (from the transferase, EC 2.7.1.37) with concomitant inactivation of the ki- rat small intestine) vanishes within a few minutes upon addition nase. This membranal proteinase exhibits a remarkable specific- of cAMP (10). The inactivation was shown to be due to the ex- ity. (i) It degrades C in its native conformation, but not after it has istence in these membranes of an enzyme that brings about a been heat-denatured. (ii) The degradation of C (Mr 40,000) does specific, limited degradation of the catalytic subunit of not proceed further, once a distinct clipped product (Mr 34,000) did not attack (under is formed. (iii) The undissociated ("stored") form of the enzyme cAMPdPKase. This membranal enzyme (R2C2) is not attacked by the membranal proteinase, preserving the same conditions) other proteins in the membrane prepa- both its potential catalytic activity and its molecular integrity.
    [Show full text]