DOCSLIB.ORG

A Recombinant System to Model Proteoglycan Aggregate

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Recombinant System to Model Proteoglycan Aggregate A RECOMBINANT SYSTEM TO MODEL PROTEOGLYCAN AGGREGATE INTERACTIONS AND AGGRECAN DEGRADATION by HAZUKI ELEANOR MIWA Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Dissertation Advisors: Dr. Thomas A. Gerken and Dr. Thomas M. Hering Department of Biochemistry CASE WESTERN RESERVE UNIVERSITY January 2006 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the dissertation of ______________________________________________________ candidate for the Ph.D. degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Copyright © 2005 by Hazuki Eleanor Miwa All rights reserved Dedications I would like to dedicate this thesis to my parents, Johji and Satsuki Bernice Miwa and my daughter, Miko Nina Miwa. Table of Contents Title page Committee Sign-off Sheet Copyright page CWRU Waiver Dedication page Table of Contents vi List of Tables xv List of Figures xvi Acknowledgements xxi List of Abbreviations xxiii Abstract xxix Chapter 1. Introduction and Background 1.1 General introduction 1.1.1 Cartilage and osteoarthritis 1 1.2 Aggregate interactions 1.2.1 Structure and function of proteoglycan aggregates in the cartilage extracellular matrix 4 1.2.2 Functions of the link protein family members 6 1.2.3 Divalent cation binding properties of link protein 8 1.2.4 Structural organization of aggrecan 9 - vi - 1.2.5 Structure and biosynthesis of KS and CS on aggrecan 12 1.2.6 Structural characteristics of link protein and the homologous G1 domain of aggrecan and other lecticans 14 1.2.7 HA binding site of PTR domain 19 1.3 Aggrecan degradation 1.3.1 Aggrecan catabolism by members of ADAMTS family 21 1.3.2 Neo-epitope antibodies 24 1.3.3 Regulation of ADAMTS4 activity 28 1.3.4 Aggrecan structure and substrate specificity of ADAMTS4 31 1.4 Focus of thesis work 36 Part I. Characterization of Recombinant Link Protein and Recombinant Aggrecan Chapter 2. Expression, Purification, and Refolding of a Pair of Recombinant Proteoglycan Tandem Repeat Domains of Link Protein from Escherichia coli Summary 38 2.1 Introduction 39 2.2 Results and Discussion 2.2.1 Cloning of link protein fragments from bovine and human link protein 41 2.2.2 Expression and purification of MBP/full-length and truncated recombinant bovine and human link protein fusion proteins in E. coli 45 2.2.3 Factor Xa digestion of MBP-bovine PTR1+2 fusion protein 49 2.2.4 Factor Xa digestion of MBP-human link proteins 54 2.2.5 Enterokinase digestion of MBP/E-bPTR1+2 56 - vii - 2.2.6 Refolding of monomeric PTR1+2 domains 58 2.2.7 Zinc (II) binding of various link protein constructs 62 2.2.8 Alternative approaches for expressing recombinant PTR1+2 domains 65 2.3. Conclusions 75 2.4. Experimental Procedures 2.4.1 Materials 76 2.4.2 Construction of E. coli link protein expression vectors 77 2.4.3 Expression and purification of MBP/full-length and truncated recombinant bovine and human link protein fusion proteins in E. coli 80 2.4.4. Factor Xa digestion of MBP fusion link proteins 82 2.4.5 SDS-PAGE and Western blot analysis 82 2.4.6 Enterokinase digestion of MBP fusion link protein 83 2.4.7 Sephacryl S-300 chromatography and Refolding of MBP/bPTR1+2 84 2.4.8 Zinc binding analysis of MBP/rhLP fusion proteins expressed in E. coli 85 2.4.9 Enzyme linked immunosorbent assay (ELISA) 86 2.4.10 Construction of bovine PTR1+2 into the pBAD/Thio-TOPO vector 86 2.4.11 Pilot expression of thioredoxin fusion PTR1+2 domains in E. coli 87 2.4.12 Construction of bovine PTR1+2 into the pPICZα vector 88 2.4.13 Pilot expression of PTR1+2 domains in Pichia pastoris 89 2.4.14 Large-scale expression and purification of the PTR1+2 domains in Pichia pastoris 90 2.4.15 N-terminal amino acid sequencing 91 - viii - Acknowledgements 92 Chapter 3. Biochemical and Functional Characterization of Full-length Recombinant Aggrecan and Cartilage Link Protein Expressed in Mammalian Cells Summary 93 3.1 Introduction 94 3.2 Results 3.2.1 Construction of recombinant aggrecan expression vectors 95 3.2.2 Mammalian expression of recombinant aggrecan 97 3.2.3 Hydrodynamic sizes of recombinant aggrecan monomer and attached glycosaminoglycans 106 3.2.4 Expression of recombinant link protein 108 3.2.5 Zinc binding of human recombinant link protein 115 3.2.6 Recombinant proteoglycan ternary aggregate formation 116 3.3 Discussion 121 3.4 Experimental Procedures 3.4.1 Materials 128 3.4.2 Purification of aggrecan and link protein from bovine cartilage and chondrocyte cultures 129 3.4.3 Construction of full-size aggrecan expression vectors 132 3.4.4 Construction of link protein expression vectors 134 3.4.5 Cell culture 135 3.4.6 Expression and purification of recombinant aggrecan in mammalian cells 136 - ix - 3.4.7 Composite agarose polyacrylamide gel analysis and chemiluminescent Western blot analysis of aggrecan 137 3.4.8 3.0% and 3.5 % SDS-PAGE gel analysis of full-sized recombinant aggrecan 138 3.4.9 Aggrecan monomer size determination by Sepharose CL-2B chromatography139 3.4.10 GAG size determination by Sepharose CL-6B chromatography 139 3.4.11 Expression and purification of recombinant bovine link protein 140 3.4.12 PNGase F digestion of recombinant link protein 141 3.4.13 Biotin labeled HA binding of link protein 142 3.4.14 Trypsin digestion of proteoglycan aggregates 142 3.4.15 Zinc (II) binding analysis of human recombinant link protein 143 3.4.16 Analysis of proteoglycan aggregate formation 144 Acknowledgements 145 Part II. Aggrecan Degradation Chapter 4. Characterization of the Substrate Specificity of ADAMTS4 against Aggrecan Core Protein Summary 146 4.1 Introduction 149 4.2 Results 4.2.1 Aggrecan structure and aggrecan catabolites 151 4.2.2 Characterization of ADAMTS4-p68 154 4.2.3 Substrate specificity of p68 and p40 156 - x - 4.2.4 KS and CS affect substrate specificity of ADAMTS4 159 4.2.5 N-linked oligosaccharides inhibit the cleavage within the IGD by ADAMTS4 165 4.2.6 Characterization of FLAG-tagged full-length aggrecan 169 4.2.7 Representative digestion of wild-type FLAG-rbAgg expressed in COS-7 cells with ADAMTS4 173 4.2.8 Anti-NITEGE reactive fragments have intact FLAG epitope and are differentially glycosylated 175 4.2.9 KS is not required for ADAMTS4 cleavage within the IGD of FLAG-rbAgg 177 4.2.10 Substrate specificity of ADAMTS4 on chondroitin sulfate-free FLAG-aggrecan. 179 4.2.11 Construction of mutagenized full-length bovine aggrecan expression vectors. 185 4.2.12 Expression of full-length FLAG-rbAgg mutant aggrecans and their susceptibility to ADAMTS4-p40 191 4.2.13 Substrate specificity of ADAMTS4-p68 on mutant aggrecans lacking potentially glycosylated residues 197 4.2.14 The role of the extended structure N-terminal to the ADAMTS4 cleavage site within the IGD. 200 4.2.15 Representative digestion of wild-type FLAG-rbAgg expressed in COS-7 cells with MMP13 205 4.2.16 MMP13 digestion of mutant aggrecans 206 - xi - 4.2.17 Keratan sulfate synthesis in COS-7, CHO-K1, and RCS cells 210 4.2.18 Co-expression of sulfotransferase and FLAG-rbAgg construct 214 4.2.19 Susceptibility of KS-FLAG-rbAgg to ADAMTS4 216 4.3 Discussion 220 4.3.1 Effects of chondroitin sulfate and keratan sulfate on substrate specificity of ADAMTS4 220 4.3.2 Substrate specificity of ADAMTS4-p68 222 4.3.3 Substrate specificity of ADAMTS4-p40 224 4.3.4 Effects of Keratan sulfate substitution on cleavage within the IGD 225 4.3.5 Effects of non-GAG oligosaccharides on aggrecan cleavage by ADAMTS4 227 4.3.6 S377 is important for substrate recognition by ADAMTS4 228 4.3.7 The potential role of the T352IQTVT357 sequence on ADAMTS4 cleavage of aggrecan 229 4.3.8 Sulfation of CS by KS sulfotransferases? 232 4.3.9 The susceptibility of KS-FLAG-rbAgg to ADAMTS4 234 4.3.10 MMP13 cleavage sites within the aggrecan core protein 236 4.3.11 Conclusions 237 4.4 Experimental Procedures 4.4.1 Materials 239 4.4.2 Site-directed mutagenesis 240 4.4.3 ADAMTS4 digestion of de-glycosylated cartilage-derived steer aggrecan 241 4.4.4 Removal of N-linked oligosaccharides from cartilage-derived aggrecan 241 - xii - 4.4.5 Secondary structure prediction 242 4.4.6 ADAMTS4 digestion of recombinant aggrecan (FLAG-rbAgg) 242 4.4.7 MMP13 digestion of recombinant aggrecan (FLAG-rbAgg) 243 4.4.8 3.0 % SDS-PAGE of full-size FLAG-rbAgg 244 4.4.9 Western blot analysis 244 4.4.10 Semi-quantification of enzymatically cleaved products 245 4.4.11 Immunocytochemistry 245 4.4.12 Expression of keratan sulfate in COS-7, CHO, and RCS cells 246 4.4.13 Co-expression of FLAG-rbAgg with sulfo- and glycosyl- transferases 247 Acknowledgements 248 Chapter 5. Summary and Future Studies 5.1 General summary 249 5.2 Future studies -Proteoglycan aggregate interactions- 5.2.1 Refolding PTR1+2 domains from E. coli 252 5.2.2 Functional characterization of the cartilage link protein-HA interaction 253 5.2.3 Effect of glycosylation on HA binding of the G1 domain of aggrecan and link protein 257 -Aggrecan degradation- 5.2.4 The presence of chondroitin sulfate and keratan sulfate on aggrecan core protein affects the substrate specificity of ADAMTS4 isoforms 259 5.2.5 Characterization of glycosylation in the IGD of recombinant aggrecan expressed - xiii - in COS-7 cells 260 5.2.6 The role of Ser 377 in ADAMTS4 recognition 261 5.2.7.
Load more

Recommended publications
  • Distribution and Clinical Significance of Heparan Sulfate Proteoglycans in Ovarian Cancer
    5178 Vol. 10, 5178–5186, August 1, 2004 Clinical Cancer Research Distribution and Clinical Significance of Heparan Sulfate Proteoglycans in Ovarian Cancer E. June Davies,1 Fiona H. Blackhall,1 decan-1 and glypican-1 were poor prognostic factors for Jonathan H. Shanks,2 Guido David,3 survival in univariate analysis. Alan T. McGown,4 Ric Swindell,5 Conclusion: We report for the first time distinct pat- 6 7 terns of expression of cell surface and extracellular matrix Richard J. Slade, Pierre Martin-Hirsch, heparan sulfate proteoglycans in normal ovary compared 1 1 John T. Gallagher, and Gordon C. Jayson with ovarian tumors. These data reinforce the role of the 1Cancer Research UK and University of Manchester Department of tumor stroma in ovarian adenocarcinoma and suggest that Medical Oncology, Paterson Institute for Cancer Research, stromal induction of syndecan-1 contributes to the patho- Manchester, England; 2Department of Histopathology, Christie Hospital NHS Trust, Manchester, England; 3Department of Medicine, genesis of this malignancy. University of Leuven, Leuven, Belgium.; 4Cancer Research UK Department of Experimental Pharmacology, Paterson Institute for Cancer Research, Manchester, England; 5Department of Medical INTRODUCTION Statistics, Christie Hospital NHS Trust, Manchester, England; The heparan sulfate proteoglycans (HSPGs) play diverse 6Department of Obstetrics and Gynaecology, Hope Hospital, Salford, Manchester, England; 7Department of Gynaecological Oncology, St. roles in tumor biology by mediating adhesion and migration
    [Show full text]
  • Versican V2 Assembles the Extracellular Matrix Surrounding the Nodes of Ranvier in the CNS
    The Journal of Neuroscience, June 17, 2009 • 29(24):7731–7742 • 7731 Cellular/Molecular Versican V2 Assembles the Extracellular Matrix Surrounding the Nodes of Ranvier in the CNS María T. Dours-Zimmermann,1 Konrad Maurer,2 Uwe Rauch,3 Wilhelm Stoffel,4 Reinhard Fa¨ssler,5 and Dieter R. Zimmermann1 Institutes of 1Surgical Pathology and 2Anesthesiology, University Hospital Zurich, CH-8091 Zurich, Switzerland, 3Vascular Wall Biology, Department of Experimental Medical Science, University of Lund, S-221 00 Lund, Sweden, 4Center for Biochemistry, Medical Faculty, University of Cologne, D-50931 Cologne, Germany, and 5Department of Molecular Medicine, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany The CNS-restricted versican splice-variant V2 is a large chondroitin sulfate proteoglycan incorporated in the extracellular matrix sur- rounding myelinated fibers and particularly accumulating at nodes of Ranvier. In vitro, it is a potent inhibitor of axonal growth and therefore considered to participate in the reduction of structural plasticity connected to myelination. To study the role of versican V2 during postnatal development, we designed a novel isoform-specific gene inactivation approach circumventing early embryonic lethality of the complete knock-out and preventing compensation by the remaining versican splice variants. These mice are viable and fertile; however, they display major molecular alterations at the nodes of Ranvier. While the clustering of nodal sodium channels and paranodal structures appear in versican V2-deficient mice unaffected, the formation of the extracellular matrix surrounding the nodes is largely impaired. The conjoint loss of tenascin-R and phosphacan from the perinodal matrix provide strong evidence that versican V2, possibly controlled by a nodal receptor, organizes the extracellular matrix assembly in vivo.
    [Show full text]
  • What Are the Roles of Metalloproteinases in Cartilage and Bone Damage? G Murphy, M H Lee
    iv44 Ann Rheum Dis: first published as 10.1136/ard.2005.042465 on 20 October 2005. Downloaded from REPORT What are the roles of metalloproteinases in cartilage and bone damage? G Murphy, M H Lee ............................................................................................................................... Ann Rheum Dis 2005;64:iv44–iv47. doi: 10.1136/ard.2005.042465 enzyme moiety into an upper and a lower subdomain. A A role for metalloproteinases in the pathological destruction common five stranded beta-sheet and two alpha-helices are in diseases such as rheumatoid arthritis and osteoarthritis, always found in the upper subdomain with a further C- and the irreversible nature of the ensuing cartilage and bone terminal helix in the lower subdomain. The catalytic sites of damage, have been the focus of much investigation for the metalloproteinases, especially the MMPs, have been several decades. This has led to the development of broad targeted for the development of low molecular weight spectrum metalloproteinase inhibitors as potential therapeu- synthetic inhibitors with a zinc chelating moiety. Inhibitors tics. More recently it has been appreciated that several able to fully differentiate between individual enzymes have families of zinc dependent proteinases play significant and not been identified thus far, although a reasonable level of varied roles in the biology of the resident cells in these tissues, discrimination is now being achieved in some cases.7 Each orchestrating development, remodelling, and subsequent family does, however, have other unique domains with pathological processes. They also play key roles in the numerous roles, including the determination of physiological activity of inflammatory cells. The task of elucidating the substrate specificity, ECM, or cell surface localisation (fig 1).
    [Show full text]
  • Quantikine® ELISA
    Quantikine® ELISA Human ADAMTS13 Immunoassay Catalog Number DADT130 For the quantitative determination of human A Disintegrin And Metalloproteinase with Thombospondin type 1 motif, 13 (ADAMTS13) concentrations in cell culture supernates, serum, and plasma. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures. TABLE OF CONTENTS SECTION PAGE INTRODUCTION .....................................................................................................................................................................1 PRINCIPLE OF THE ASSAY ...................................................................................................................................................2 LIMITATIONS OF THE PROCEDURE .................................................................................................................................2 TECHNICAL HINTS .................................................................................................................................................................2 MATERIALS PROVIDED & STORAGE CONDITIONS ...................................................................................................3 OTHER SUPPLIES REQUIRED .............................................................................................................................................4 PRECAUTIONS .........................................................................................................................................................................4
    [Show full text]
  • Kids First Pediatric Research Program (Kids First) Poster Session at ASHG Accelerating Pediatric Genomics Research Through Collaboration October 15Th, 2019
    The Gabriella Miller Kids First Pediatric Research Program (Kids First) Poster Session at ASHG Accelerating Pediatric Genomics Research through Collaboration October 15th, 2019 Background The Gabriella Miller Kids First Pediatric Research Program (Kids First) is a trans- NIH Common Fund program initiated in response to the 2014 Gabriella Miller Kids First Research Act. The program’s vision is to alleviate suffering from childhood cancer and structural birth defects by fostering collaborative research to uncover the etiology of these diseases and support data sharing within the pediatric research community. This is implemented through developing the Gabriella Miller Kids First Data Resource (Kids First Data Resource) and populating this resource with whole genome sequence datasets and associated clinical and phenotypic information. Both childhood cancers and structural birth defects are critical and costly conditions associated with substantial morbidity and mortality. Elucidating the underlying genetic etiology of these diseases has the potential to profoundly improve preventative measures, diagnostics, and therapeutic interventions. Purpose During this evening poster session, attendees will gain a broad understanding of the utility of the genomic data generated by Kids First, learn about the progress of Kids First X01 cohort projects, and observe demonstrations of the tools and functionalities of the recently launched Kids First Data Resource Portal. The session is an opportunity for the scientific community and public to engage with Kids First investigators, collaborators, and a growing community of researchers, patient foundations, and families. Several other NIH and external data efforts will present posters and be available to discuss collaboration opportunities as we work together to accelerate pediatric research.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • ADAMTS13 and 15 Are Not Regulated by the Full Length and N‑Terminal Domain Forms of TIMP‑1, ‑2, ‑3 and ‑4
    BIOMEDICAL REPORTS 4: 73-78, 2016 ADAMTS13 and 15 are not regulated by the full length and N‑terminal domain forms of TIMP‑1, ‑2, ‑3 and ‑4 CENQI GUO, ANASTASIA TSIGKOU and MENG HUEE LEE Department of Biological Sciences, Xian Jiaotong-Liverpool University, Suzhou, Jiangsu 215123, P.R. China Received June 29, 2015; Accepted July 15, 2015 DOI: 10.3892/br.2015.535 Abstract. A disintegrin and metalloproteinase with thom- proteolysis activities associated with arthritis, morphogenesis, bospondin motifs (ADAMTS) 13 and 15 are secreted zinc angiogenesis and even ovulation [as reviewed previously (1,2)]. proteinases involved in the turnover of von Willebrand factor Also known as the VWF-cleaving protease, ADAMTS13 and cancer suppression. In the present study, ADAMTS13 is noted for its ability in cleaving and reducing the size of the and 15 were subjected to inhibition studies with the full-length ultra-large (UL) form of the VWF. Reduction in ADAMTS13 and N-terminal domain forms of tissue inhibitor of metallo- activity from either hereditary or acquired deficiency causes proteinases (TIMPs)-1 to -4. TIMPs have no ability to inhibit accumulation of UL-VWF multimers, platelet aggregation and the ADAMTS proteinases in the full-length or N-terminal arterial thrombosis that leads to fatal thrombotic thrombocy- domain form. While ADAMTS13 is also not sensitive to the topenic purpura [as reviewed previously (1,3)]. By contrast, hydroxamate inhibitors, batimastat and ilomastat, ADAMTS15 ADAMTS15 is a potential tumor suppressor. Only a limited app can be effectively inhibited by batimastat (Ki 299 nM). In number of in-depth investigations have been carried out on the conclusion, the present results indicate that TIMPs are not the enzyme; however, expression and profiling studies have shown regulators of these two ADAMTS proteinases.
    [Show full text]
  • ADAMTS Proteases in Vascular Biology
    Review MATBIO-1141; No. of pages: 8; 4C: 3, 6 ADAMTS proteases in vascular biology Juan Carlos Rodríguez-Manzaneque 1, Rubén Fernández-Rodríguez 1, Francisco Javier Rodríguez-Baena 1 and M. Luisa Iruela-Arispe 2 1 - GENYO, Centre for Genomics and Oncological Research, Pfizer, Universidad de Granada, Junta de Andalucía, 18016 Granada, Spain 2 - Department of Molecular, Cell, and Developmental Biology, Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA Correspondence to Juan Carlos Rodríguez-Manzaneque and M. Luisa Iruela-Arispe: J.C Rodríguez-Manzaneque is to be contacted at: GENYO, 15 PTS Granada - Avda. de la Ilustración 114, Granada 18016, Spain; M.L. Iruela-Arispe, Department of Molecular, Cell and Developmental Biology, UCLA, 615 Charles Young Drive East, Los Angeles, CA 90095, USA. [email protected]; [email protected] http://dx.doi.org/10.1016/j.matbio.2015.02.004 Edited by W.C. Parks and S. Apte Abstract ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) proteases comprise the most recently discovered branch of the extracellular metalloenzymes. Research during the last 15 years, uncovered their association with a variety of physiological and pathological processes including blood coagulation, tissue repair, fertility, arthritis and cancer. Importantly, a frequent feature of ADAMTS enzymes relates to their effects on vascular-related phenomena, including angiogenesis. Their specific roles in vascular biology have been clarified by information on their expression profiles and substrate specificity. Through their catalytic activity, ADAMTS proteases modify rather than degrade extracellular proteins. They predominantly target proteoglycans and glycoproteins abundant in the basement membrane, therefore their broad contributions to the vasculature should not come as a surprise.
    [Show full text]
  • Three Is a Crowd
    TREPAR 1600 No. of Pages 12 Review Three Rosetting[403_TD$IF] in Plasmodium falciparum Xue Yan Yam,1,3 Makhtar Niang,2,3 Kripa Gopal Madnani,1,3 and Peter R. Preiser1,* The intracellular malaria parasites extensively modify host erythrocytes to allow Trends nutrient uptake, ensure homeostasis, and evade the host’s immune response. Rosetting, the binding of uninfected To achieve this, the parasite exports several proteins to the erythrocyte surface. RBCs to a parasite-infected RBC In Plasmodium falciparum, the parasite responsible for the most severe form of (iRBC), has been directly linked to human malaria, three major variant surface antigen families – PfEMP1, STE- the severity of clinical disease. VOR, and RIFIN – have been implicated in contributing to immune evasion, Three parasite protein families, parasite sequestration, and parasite-mediated rosetting of uninfected eryth- PfEMP1, STEVOR, and RIFIN, mediate rosetting in Plasmodium falciparum. rocytes. Sequestration and rosetting have been linked to parasite-mediated pathology, making the variant surface antigens of P. falciparum major virulence Sequential timing of surface expres- factors. Here we review our current understanding of rosetting mechanism, sion of PfEMP1, RIFIN, and STEVOR fi on iRBC suggests that the parasite has recent ndings of STEVOR, RIFIN-mediated rosetting, and their implication on developed three different rosette for- the severity and pathology of the disease. mation mechanisms, implicating a cri- tical function for parasite survival. Plasmodium Variant Surface Antigens of Parasites: A Role in Rosetting PfEMP1 mediates rosetting through Plasmodium parasites have a complex life cycle involving a mosquito vector and a mammalian CR1, heparan sulfate, and blood group host.
    [Show full text]
  • TACI:Fc Scavenging B Cell Activating Factor (BAFF) Alleviates Ovalbumin-Induced Bronchial Asthma in Mice
    EXPERIMENTAL and MOLECULAR MEDICINE, Vol. 39, No. 3, 343-352, June 2007 TACI:Fc scavenging B cell activating factor (BAFF) alleviates ovalbumin-induced bronchial asthma in mice 1,2,3 2 Eun-Yi Moon and Sook-Kyung Ryu the percentage of non-lymphoid cells and no changes were detected in lymphoid cell population. 1 Department of Bioscience and Biotechnology Hypodiploid cell formation in BALF was decreased Sejong University by OVA-challenge but it was recovered by TACI:Fc Seoul 143-747, Korea treatment. Collectively, data suggest that asthmatic 2 Laboratory of Human Genomics symptom could be alleviated by scavenging BAFF Korea Research Institute of Bioscience and Biotechnology (KRIBB) and then BAFF could be a novel target for the Daejeon 305-806, Korea develpoment of anti-asthmatic agents. 3 Corresponding author: Tel, 82-2-3408-3768; Fax, 82-2-466-8768; E-mail, [email protected] Keywords: asthma; B-cell activating factor; ovalbu- and [email protected] min; transmembrane activator and CAML interactor protein Accepted 28 March 2007 Introduction Abbreviations: BAFF, B cell activating factor belonging to TNF- family; BALF, bronchoalveolar lavage fluid; OVA, ovalbumin; PAS, Mature B cell generation and maintenance are regu- periodic acid-Schiff; Prx, peroxiredoxin; TACI, transmembrane lated by B-cell activating factor (BAFF). BAFF is pro- activator and calcium modulator and cyclophilin ligand interactor duced by macrophages or dendritic cells upon stim- ulation with LPS or IFN- . BAFF belongs to the TNF family. Its biological role is mediated by the specific Abstract receptors, B-cell maturation antigen (BCMA), trans- membrane activator and calcium modulator and cy- Asthma was induced by the sensitization and chal- clophilin ligand interactor (TACI) and BAFF receptor, lenge with ovalbumin (OVA) in mice.
    [Show full text]
  • A Single-Cell Transcriptomic Landscape of Primate Arterial Aging
    ARTICLE https://doi.org/10.1038/s41467-020-15997-0 OPEN A single-cell transcriptomic landscape of primate arterial aging Weiqi Zhang 1,2,3,4,5,13, Shu Zhang6,7,13, Pengze Yan3,8,13, Jie Ren7,9,13, Moshi Song3,5,8, Jingyi Li2,3,8, Jinghui Lei4, Huize Pan2,3, Si Wang3,5,8, Xibo Ma3,10, Shuai Ma2,3,8, Hongyu Li2,3, Fei Sun2,3, Haifeng Wan3,5,11, ✉ ✉ ✉ Wei Li 3,5,11, Piu Chan4, Qi Zhou3,5,11, Guang-Hui Liu 2,3,4,5,8 , Fuchou Tang 6,7,9,12 & Jing Qu 3,5,11 Our understanding of how aging affects the cellular and molecular components of the vas- 1234567890():,; culature and contributes to cardiovascular diseases is still limited. Here we report a single-cell transcriptomic survey of aortas and coronary arteries in young and old cynomolgus monkeys. Our data define the molecular signatures of specialized arteries and identify eight markers discriminating aortic and coronary vasculatures. Gene network analyses characterize tran- scriptional landmarks that regulate vascular senility and position FOXO3A, a longevity- associated transcription factor, as a master regulator gene that is downregulated in six subtypes of monkey vascular cells during aging. Targeted inactivation of FOXO3A in human vascular endothelial cells recapitulates the major phenotypic defects observed in aged monkey arteries, verifying FOXO3A loss as a key driver for arterial endothelial aging. Our study provides a critical resource for understanding the principles underlying primate arterial aging and contributes important clues to future treatment of age-associated vascular disorders. 1 CAS Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]