Distinct Biological Events Generated by ECM Proteolysis by Two Homologous Collagenases

Total Page:16

File Type:pdf, Size:1020Kb

Distinct Biological Events Generated by ECM Proteolysis by Two Homologous Collagenases Distinct biological events generated by ECM proteolysis by two homologous collagenases Inna Solomonova, Eldar Zehoraia, Dalit Talmi-Franka,b, Sharon G. Wolfc, Alla Shainskayad, Alina Zhuravleva, Elena Kartvelishvilyc, Robert Vissee, Yishai Levind, Nir Kampff, Diego Adhemar Jaitinb, Eyal Davidb, Ido Amitb, Hideaki Nagasee, and Irit Sagia,1 aDepartment of Biological Regulation, Weizmann Institute of Science, Rehovot 76100, Israel; bDepartment of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel; cDepartment of Chemical Research Support, Weizmann Institute of Science, Rehovot 76100, Israel; dNancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science, Rehovot 76100, Israel; eKennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology, and Musculoskeletal Sciences, University of Oxford, Oxford, OX3 7FY, United Kingdom; and fDepartment of Materials and Interfaces, Weizmann Institute of Science, Rehovot 76100, Israel Edited by Christopher M. Overall, University of British Columbia, Vancouver, Canada, and accepted by Editorial Board Member Rakesh K. Jain August 8, 2016 (received for review October 4, 2015) It is well established that the expression profiles of multiple and to unique ECM–cell crosstalk. We reveal that MMP-1 and MMP-13 possibly redundant matrix-remodeling proteases (e.g., collagenases) cause distinct ECM degradation, bringing about significantly distinct differ strongly in health, disease, and development. Although cellular phenotypes. Our findings show the complexity and selectivity enzymatic redundancy might be inferred from their close similarity in of collagenase-associated degradation mechanisms during tissue structure, their in vivo activity can lead to extremely diverse tissue- remodeling; these mechanisms could be used as a tool for future remodeling outcomes. We observed that proteolysis of collagen-rich therapeutic interventions. natural extracellular matrix (ECM), performed uniquely by individual homologous proteases, leads to distinct events that eventually affect Results overall ECM morphology, viscoelastic properties, and molecular Selective Degradation of ECM by Collagenases Determines Fibroblast composition. We revealed striking differences in the motility and Behavior. We set out to characterize the specific influences of the signaling patterns, morphology, and gene-expression profiles of cells – highly abundant collagenases on fibroblast ECM crosstalk. In this BIOCHEMISTRY interacting with natural collagen-rich ECM degraded by different study we used natural collagen fascicles (termed “ECM”)from collagenases. Thus, in contrast to previous notions, matrix-remodeling tendons of 6-mo-old rats (22, 23). We examined the effect of this systems are not redundant and give rise to precise ECM–cell crosstalk. ECMonrat-1fibroblasts,whichareknowntobesensitivetoECM Because ECM proteolysis is an abundant biochemical process that is composition and rigidity. Moreover, these cells, much like epithe- critical for tissue homoeostasis, these results improve our fundamen- lial cells, can exhibit different modes of migration toward different tal understanding its complexity and its impact on cell behavior. substrates and engage in cell–cell interactions (24, 25). Because fibroblasts inherently express ECM proteins and remodeling en- ECM | MMP | proteolysis zymes, we conducted our experiments at early stages of interaction he function and integrity of the extracellular matrix (ECM) is Significance Tvital for cell behavior as well as for whole-tissue homeostasis – (1 4). The ECM undergoes constant remodeling during health Extracellular matrix (ECM) proteolysis is an abundant bio- and disease states. Its components are regularly being deposited, chemical process. Here we describe the multilayered biological degraded, or otherwise modified. The highly stable fibrillar col- complexity generated by structurally homologous collagenases lagen type I (Col I) is abundant in many organ-derived ECMs (matrix metalloproteinase-1 and matrix metalloproteinase-13) in and connective tissues (5, 6); it serves as a tissue scaffold, deter- collagen-rich, native ECM, that may prove central to tissue ho- mining ECM mechanical properties and anchoring other ECM meostasis and pathology. The events induced by these two col- proteins necessary for cell function (7). These processes are or- lagenases generate microenvironments characterized by distinct chestrated by multiple remodeling enzymes, among which the ma- chemical, biomechanical, and morphological ECM properties that trix metalloproteinase (MMP) family plays an important role. Only lead to different cellular behaviors. Our findings improve the a few members of this proteinase family, the collagenases, are able fundamental understanding of selective ECM degradation by to degrade the resistant fibrillar collagens, i.e., Col I, as well as other homologous collagenases and its impact on cell behavior. ECM molecules (8, 9). The collagenases have conserved amino acids in their zinc-containing catalytic domain (8, 10) and display Author contributions: I. Solomonov, E.Z., I.A., H.N., and I. Sagi designed research; I. Solomonov, high structural similarities, as reflected by their functional domain E.Z., S.G.W., A.S., A.Z., E.K., Y.L., N.K., and D.A.J. performed research; R.V. and H.N. contrib- organization. Nevertheless, the complex effects exerted by different uted new reagents/analytic tools; I. Solomonov, E.Z., D.T.-F., A.S., Y.L., D.A.J., E.D., and MMPs on ECM and cells in vivo remain poorly understood. I. Sagi analyzed data; and I. Solomonov, E.Z., D.T.-F., S.G.W., and I. Sagi wrote the paper. The enzymatic activity of MMPs, and specifically of collage- The authors declare no conflict of interest. nases, in vivo is tightly regulated (11), with enzymatic dysregu- This article is a PNAS Direct Submission. C.M.O. is a Guest Editor invited by the Editorial lation causing irreversible damage associated with a variety of Board. diseases. Abnormally elevated levels of MMP-1 or of both MMP-1 Data deposition: MS data and protein/peptide identifications from in-gel analysis have been uploaded to the ProteomeXchange Consortium via the PRIDE repository, www.ebi. and MMP-13 have been associated with different types of cancers ac.uk/pride/archive/login (project accession no. PXD003796; username: reviewer59836@ andwithinflammatorydiseases(12–21). ebi.ac.uk; password: sTHj8wE8). Mass spectrometry data and protein/peptide identifica- Here, we explored the degradation patterns of natural collagen- tions from in-solution digestion have been uploaded to the ProteomeXchange Consor- rich ECM by two homologous MMPs and tested the response of tium via the PRIDE repository, www.ebi.ac.uk/pride/archive/login (project accession no. PXD003553; username: [email protected]; password: cJGfrm0Z). The RNA- cells to intact vs. selectively degraded ECM. We collectively pro- sequencing dataset has been deposited in the Gene Expression Omnibus database (acces- filed the unique remodeling events caused by two secreted colla- sion no. GSE79749). genases, MMP-1 and MMP-13, by using biochemical, biophysical, 1To whom correspondence should be addressed. Email: [email protected]. and proteomics tools. We show that these proteases drive morpho- This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. logical, biochemical, and viscoelastic changes in the ECM leading 1073/pnas.1519676113/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1519676113 PNAS Early Edition | 1of6 Downloaded by guest on September 23, 2021 (up to 4 h); in this time frame, no collagen deposition or MMP-1 adopted round, symmetrical morphologies. To validate these and MMP-13 secretion was detected (Fig. S1 A and B). morphological changes, we further quantified the aspect ratios of Cell motility was characterized using real-time optical imaging. cells that adhered to differentially degraded ECM using our Although fibroblasts demonstrated a clustered, directional motil- SEM images at 4 h after seeding (Fig. 1 D–F). This analysis ity toward native ECM (control) with a measured velocity of 40 ± yielded a trend in cell aspect ratio similar to that obtained with 14 μm/h, they showed individual modes of motility with a velocity live-cell imaging at the measured time point (Fig. 1H). Overall, of 37 ± 5 μm/h when moving toward MMP-1–degraded ECM. In these analyses provide cell-shape statistics indicating that rat-1 the presence of MMP-13–degraded ECM, the cells exhibited re- cells adopt different morphologies upon interaction with differ- duced or arrested motility with a measured velocity of 2 ± 2 μm/h entially degraded Col I-rich ECM. (Fig. 1 A–C, Fig. S1C,andMovies S1–S3). The differential phenotypes acquired by fibroblasts interacting To analyze the overall shape of the cells, we quantified the cell with degraded ECM suggested a typical and unique cellular re- aspect ratio (the length divided by the width) as a function of sponse driven by signal modulations transmitted by the ECM time using our live-cell imaging data (Fig. 1G). This analysis (26, 27). Because cells regulate migration, proliferation, and allowed us to compare precisely the shape of cells migrating on adhesion mainly through the activation of the ERK1/2 cascade differentially remodeled collagen-rich ECM. Cells imaged on (28), we examined the effect of ECM remodeling on this cas- control ECM showed aspect ratios ranging from 1.36 to 1.42, cade. Fibroblasts adhering to native
Recommended publications
  • MATRIX METALLOPROTEINASE REQUIREMENTS for NEUROMUSCULAR SYNAPTOGENESIS by Mary Lynn Dear Dissertation Submitted to the Faculty O
    MATRIX METALLOPROTEINASE REQUIREMENTS FOR NEUROMUSCULAR SYNAPTOGENESIS By Mary Lynn Dear Dissertation Submitted to the Faculty of the Graduate School of Vanderbilt University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biological Sciences January 31, 2018 Nashville, Tennessee Approved: Todd Graham, Ph.D. Kendal Broadie, Ph.D. Katherine Friedman, Ph.D. Barbara Fingleton, Ph.D. Jared Nordman, Ph.D. Copyright © 2018 by Mary Lynn Dear All Rights Reserved ACKNOWLEDGEMENTS I would like to acknowledge my advisor, Kendal Broadie, and the entire Broadie laboratory, both past and present, for their endless support, engaging conversations, thoughtful suggestions and constant encouragement. I would also like to thank my committee members both past and present for playing such a pivotal role in my graduate career and growth as a scientist. I thank the Department of Biological Sciences for fostering my graduate education. I thank the entire protease community for their continued support, helpful suggestions and collaborative efforts that helped my project move forward. I would like to acknowledge Dr. Andrea Page-McCaw and her entire laboratory for helpful suggestions, being engaged in my studies and providing many tools that were invaluable to my project. I thank my parents, Leisa Justus and Raymond Dear, my brother Jake Dear and my best friend Jenna Kaufman. Words cannot describe how grateful I am for your support and encouragement. Most importantly, I would like to thank my husband, Jeffrey Thomas, for always believing in me, your unwavering support and for helping me endure all the ups and downs that come with graduate school.
    [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]
  • Discovery and Optimization of Selective Inhibitors of Meprin Α (Part II)
    pharmaceuticals Article Discovery and Optimization of Selective Inhibitors of Meprin α (Part II) Chao Wang 1,2, Juan Diez 3, Hajeung Park 1, Christoph Becker-Pauly 4 , Gregg B. Fields 5 , Timothy P. Spicer 1,6, Louis D. Scampavia 1,6, Dmitriy Minond 2,7 and Thomas D. Bannister 1,2,* 1 Department of Molecular Medicine, Scripps Research, Jupiter, FL 33458, USA; [email protected] (C.W.); [email protected] (H.P.); [email protected] (T.P.S.); [email protected] (L.D.S.) 2 Department of Chemistry, Scripps Research, Jupiter, FL 33458, USA; [email protected] 3 Rumbaugh-Goodwin Institute for Cancer Research, Nova Southeastern University, 3321 College Avenue, CCR r.605, Fort Lauderdale, FL 33314, USA; [email protected] 4 The Scripps Research Molecular Screening Center, Scripps Research, Jupiter, FL 33458, USA; [email protected] 5 Unit for Degradomics of the Protease Web, Institute of Biochemistry, University of Kiel, Rudolf-Höber-Str.1, 24118 Kiel, Germany; fi[email protected] 6 Department of Chemistry & Biochemistry and I-HEALTH, Florida Atlantic University, 5353 Parkside Drive, Jupiter, FL 33458, USA 7 Dr. Kiran C. Patel College of Allopathic Medicine, Nova Southeastern University, 3301 College Avenue, Fort Lauderdale, FL 33314, USA * Correspondence: [email protected] Abstract: Meprin α is a zinc metalloproteinase (metzincin) that has been implicated in multiple diseases, including fibrosis and cancers. It has proven difficult to find small molecules that are capable Citation: Wang, C.; Diez, J.; Park, H.; of selectively inhibiting meprin α, or its close relative meprin β, over numerous other metzincins Becker-Pauly, C.; Fields, G.B.; Spicer, which, if inhibited, would elicit unwanted effects.
    [Show full text]
  • ADAM17 Targets MMP-2 and MMP-9 Via EGFR-MEK-ERK Pathway Activation to Promote Prostate Cancer Cell Invasion
    1714 INTERNATIONAL JOURNAL OF ONCOLOGY 40: 1714-1724, 2012 ADAM17 targets MMP-2 and MMP-9 via EGFR-MEK-ERK pathway activation to promote prostate cancer cell invasion LI-JIE XIAO1,2*, PING LIN1*, FENG LIN1*, XIN LIU1, WEI QIN1, HAI-FENG ZOU1, LIANG GUO1, WEI LIU1, SHU-JUAN WANG1 and XIAO-GUANG YU1 1Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Harbin Medical University, 194 Xuefu Road, Harbin 150081, Heilongjiang; 2College of Life Science and Technology, Heilongjiang Bayi Agricultural University, 2 Xinyang Road, Daqing 163319, Heilongjiang, P.R. China Received September 8, 2011; Accepted November 18, 2011 DOI: 10.3892/ijo.2011.1320 Abstract. ADAM17, also known as tumor necrosis factor-α released and down-regulation of MMP-2, MMP-9. However, converting enzyme (TACE), is involved in proteolytic ectodo- these effects could be reversed by simultaneous addition of main shedding of cell surface molecules and cytokines. TGF-α. These data demonstrated that ADAM17 contributes to Although aberrant expression of ADAM17 has been shown androgen-independent prostate cancer cell invasion by shed- in various malignancies, the function of ADAM17 in prostate ding of EGFR ligand TGF-α, which subsequently activates the cancer has not been clarified. In the present study, we sought EGFR-MEK-ERK signaling pathway, leading finally to over- to elucidate whether ADAM17 contributes to prostate cancer expression of MMP-2 and MMP-9. This study suggests that the cell invasion, as well as the mechanism involved in the process. ADAM17 expression level may be a new predictive biomarker The expression pattern of ADAM17 was investigated in human of invasion and metastasis of prostate cancer, and ADAM17 prostate cancer cells.
    [Show full text]
  • On and Polymorphisms in Q Fever
    Matrix metalloproteinase expression, produc3on and polymorphisms in Q fever Anne F.M. Jansen1,2, Teske Schoffelen1,2, Julien Textoris3, Jean Louis Mege3, Chantal P. Bleeker-Rovers1,2, Esther van de Vosse4, Hendrik Jan Roest5, Marcel van Deuren1,2 1. Department of Internal Medicine, Division of Experimental Medicine, Radboud university medical center, Nijmegen, The Netherlands 2. Radboud Expert Centre for Q fever, Radboud university medical center, Nijmegen, the Netherlands, 3. URMITE, CNRS UMR 7278, IRD 198, INSERM 1095 Aix-Marseille University, Marseille, France 4. Department of Infec3ous Diseases, Leiden University Medical Center, Leiden, The Netherlands 5. Department of Bacteriology and TSEs, Central Veterinary Instute, part of Wageningen UR, Lelystad, the Netherlands Background C. burnei induces MMP-1 and MMP-9 produc3on in PBMCs Chronic Q fever is a life threatening condi3on caused by the Gram-negave bacterium Coxiella burnei, manifes3ng as an infec3on of aneurysms, aor3c prosthesis or heart valves. Matrix metalloproteinases (MMPs) are proteoly3c enzymes that cleave extracellular matrix and are implicated in the pathology of aneurysms and endocardi3s. Currently, the contribu3on of MMPs to the pathogenesis of chronic Q fever is unknown. Methods We inves3gated the C. burnei specific gene expression of MMPs in PBMCs and protein produc3on by ELISA in chronic Q fever paents (n=6, n=10, respec3vely), cardiovascular paents with a history of Q fever (n=10) and healthy controls (n=4, n=10, respec3vely), in some experiments, the controls had vascular disease (n=10). Circulang MMP levels were assessed with Luminex technology and these groups were also genotyped for 20 SNPs in MMP and Tissue Inhibitor of MMP (TIMP) genes.
    [Show full text]
  • Matrix Metalloproteinases in Angiogenesis: a Moving Target for Therapeutic Intervention
    Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention William G. Stetler-Stevenson J Clin Invest. 1999;103(9):1237-1241. https://doi.org/10.1172/JCI6870. Perspective Angiogenesis is the process in which new vessels emerge from existing endothelial lined vessels. This is distinct from the process of vasculogenesis in that the endothelial cells arise by proliferation from existing vessels rather than differentiating from stem cells. Angiogenesis is an invasive process that requires proteolysis of the extracellular matrix and, proliferation and migration of endothelial cells, as well as synthesis of new matrix components. During embryonic development, both vasculogenesis and angiogenesis contribute to formation of the circulatory system. In the adult, with the single exception of the reproductive cycle in women, angiogenesis is initiated only in response to a pathologic condition, such as inflammation or hypoxia. The angiogenic response is critical for progression of wound healing and rheumatoid arthritis. Angiogenesis is also a prerequisite for tumor growth and metastasis formation. Therefore, understanding the cellular events involved in angiogenesis and the molecular regulation of these events has enormous clinical implications. This understanding is providing novel therapeutic targets for the treatment of a variety of diseases, including cancer. Whatever the pathologic condition, an initiating stimulus results in the formation of a migrating solid column of endothelial cells called the vascular sprout. The advancing front of this endothelial cell column presumably focuses proteolytic activity to create a defect in the extracellular matrix, through which the advancing and proliferating column of endothelial […] Find the latest version: https://jci.me/6870/pdf Matrix metalloproteinases in angiogenesis: Perspective a moving target for therapeutic intervention SERIES Topics in angiogenesis David A.
    [Show full text]
  • Gent Forms of Metalloproteinases in Hydra
    Cell Research (2002); 12(3-4):163-176 http://www.cell-research.com REVIEW Structure, expression, and developmental function of early diver- gent forms of metalloproteinases in Hydra 1 2 3 4 MICHAEL P SARRAS JR , LI YAN , ALEXEY LEONTOVICH , JIN SONG ZHANG 1 Department of Anatomy and Cell Biology University of Kansas Medical Center Kansas City, Kansas 66160- 7400, USA 2 Centocor, Malvern, PA 19355, USA 3 Department of Experimental Pathology, Mayo Clinic, Rochester, MN 55904, USA 4 Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA ABSTRACT Metalloproteinases have a critical role in a broad spectrum of cellular processes ranging from the breakdown of extracellular matrix to the processing of signal transduction-related proteins. These hydro- lytic functions underlie a variety of mechanisms related to developmental processes as well as disease states. Structural analysis of metalloproteinases from both invertebrate and vertebrate species indicates that these enzymes are highly conserved and arose early during metazoan evolution. In this regard, studies from various laboratories have reported that a number of classes of metalloproteinases are found in hydra, a member of Cnidaria, the second oldest of existing animal phyla. These studies demonstrate that the hydra genome contains at least three classes of metalloproteinases to include members of the 1) astacin class, 2) matrix metalloproteinase class, and 3) neprilysin class. Functional studies indicate that these metalloproteinases play diverse and important roles in hydra morphogenesis and cell differentiation as well as specialized functions in adult polyps. This article will review the structure, expression, and function of these metalloproteinases in hydra. Key words: Hydra, metalloproteinases, development, astacin, matrix metalloproteinases, endothelin.
    [Show full text]
  • Effects of Collagen-Derived Bioactive Peptides and Natural Antioxidant
    www.nature.com/scientificreports OPEN Efects of collagen-derived bioactive peptides and natural antioxidant compounds on Received: 29 December 2017 Accepted: 19 June 2018 proliferation and matrix protein Published: xx xx xxxx synthesis by cultured normal human dermal fbroblasts Suzanne Edgar1, Blake Hopley1, Licia Genovese2, Sara Sibilla2, David Laight1 & Janis Shute1 Nutraceuticals containing collagen peptides, vitamins, minerals and antioxidants are innovative functional food supplements that have been clinically shown to have positive efects on skin hydration and elasticity in vivo. In this study, we investigated the interactions between collagen peptides (0.3–8 kDa) and other constituents present in liquid collagen-based nutraceuticals on normal primary dermal fbroblast function in a novel, physiologically relevant, cell culture model crowded with macromolecular dextran sulphate. Collagen peptides signifcantly increased fbroblast elastin synthesis, while signifcantly inhibiting release of MMP-1 and MMP-3 and elastin degradation. The positive efects of the collagen peptides on these responses and on fbroblast proliferation were enhanced in the presence of the antioxidant constituents of the products. These data provide a scientifc, cell-based, rationale for the positive efects of these collagen-based nutraceutical supplements on skin properties, suggesting that enhanced formation of stable dermal fbroblast-derived extracellular matrices may follow their oral consumption. Te biophysical properties of the skin are determined by the interactions between cells, cytokines and growth fac- tors within a network of extracellular matrix (ECM) proteins1. Te fbril-forming collagen type I is the predomi- nant collagen in the skin where it accounts for 90% of the total and plays a major role in structural organisation, integrity and strength2.
    [Show full text]
  • Prognostic Significance of MMP-1 and MMP-3 Functional Promoter Polymorphisms in Colorectal Cancer
    594 Vol. 11, 594–599, January 15, 2005 Clinical Cancer Research Prognostic Significance of MMP-1 and MMP-3 Functional Promoter Polymorphisms in Colorectal Cancer Franck Zinzindohoue´,1 Thierry Lecomte,2 INTRODUCTION Jean-Marc Ferraz,2 Anne-Marie Houllier,1 Matrix metalloproteinase (MMP) belongs to a large group Paul-Henri Cugnenc,2 Anne Berger,2 of proteases, which includes over 22 known human zinc- He´le`ne Blons,1 and Pierre Laurent-Puig1 dependent proteolytic enzymes. These are capable of breaking essentially all components of the extracellular matrix (1, 2). 1INSERM U490 Laboratoire de toxicologie Mole´culaire and 2 MMPs take part in high tissue turnover and remodeling, both Poˆle de cance´rologie Hoˆpital europe´en Georges Pompidou, Paris, France physiologic and pathologic conditions such as cancer. MMPs are also implicated in all steps of tumorogenesis, cancer invasion, and metastasis (3). Tumor invasion and metastasis formation ABSTRACT always begin with blood and lymphatic vessel infiltration. As Purpose: Matrix metalloproteinase (MMP) belongs to a these processes involve proteolysis of the extracellular matrix, large group of proteases capable of breaking essentially all MMPs are suggested to play a major role in tumor progression. components of the extracellular matrix. They are implicated Colorectal cancer is the most common malignancy of the in all steps of tumorogenesis, cancer invasion, and gastrointestinal tract. It is the third cause of cancer overall and metastasis. Among them, metalloproteinase type 1 (MMP- the second leading cause of cancer-related death in the Europe 1) is implicated in tumor invasion and metastasis in and the United States with an incidence of 300,000 new cases different types of cancers including colorectal cancer in (4).
    [Show full text]
  • Neprilysin Activity Assay Kit (MAK350)
    Neprilysin Activity Assay Kit Catalog Number MAK350 Storage Temperature –20 C TECHNICAL BULLETIN Product Description Components Neprilysin, also known as neutral endopeptidase, The kit is sufficient for 100 fluorometric assays in enkephalinase, CD10, and common acute 96 well plates. lymphoblastic leukemia antigen, is a zinc-containing transmembrane metalloproteinase. It is able to NEP Assay Buffer 40 mL hydrolyze very important endogenous peptides, such as Catalog Number MAK350A natriuretic atrial factor, enkephalins, substance P, bradykinin and amyloid (A ) peptide. Thus, NEP is a Neprilysin (Lyophilized) 1 vial potentially therapeutic target in important pathological Catalog Number MAK350B conditions such as cardiovascular disease, prostate cancer, and Alzheimer’s disease. NEP has also been NEP Substrate (in DMSO) 15 L used as a biological marker in a type of childhood Catalog Number MAK350C leukemia. The detection of NEP in endometrial stromal cells has been proposed as a helpful tool in the Abz-Standard (1 mM) 100 L diagnosis of endometriosis. NEP is currently a focus of Catalog Number MAK350D major interest in cardiovascular and neurological research. Reagents and Equipment Required but Not Provided. The Neprilysin Activity Assay Kit utilizes the ability of an Pipetting devices and accessories active NEP to cleave a synthetic substrate, o-amino- (e.g., multichannel pipettor) benzoic acid (Abz)-based peptide, to release a free White opaque flatbottom 96 well plates fluorophore. The released Abz can be easily quantified Fluorescence multiwell plate reader, capable of using a fluorescence microplate reader. The substrate 37 C temperature setting is specific to NEP and can differentiate the NEP activity Refrigerated microcentrifuge capable of from trypsin and other structurally similar zinc RCF 12,000 g metalloproteinase in biological samples such as Protease Inhibitor Cocktail (contains 80 M angiotensin converting enzymes (ACE1 and ACE2) and Aprotinin) (Catalog Number P8340) endothelin converting enzymes (ECE1 and ECE2).
    [Show full text]
  • Role of MMP-1, MMP-8 and MMP-9 Gene Polymorphisms in Preterm Birth
    Journal of Genetics (2020)99:2 Ó Indian Academy of Sciences https://doi.org/10.1007/s12041-019-1161-7 (0123456789().,-volV)(0123456789().,-volV) RESEARCH ARTICLE Role of MMP-1, MMP-8 and MMP-9 gene polymorphisms in preterm birth MONIKA PANDEY* and SHALLY AWASTHI Department of Pediatrics, King George’s Medical University, Lucknow 226 003, India *For correspondence. E-mail: [email protected]. Received 6 January 2019; revised 17 August 2019; accepted 18 September 2019 Abstract. Novel approaches to preterm births are underway building upon our prior discoveries and probing into unknown discovery pathways. The recent findings showed a high affinity of MMP-9 in serum and its polymorphisms for preterm birth. This study, which is a hospital-based case– control study, aims to investigate the association of MMP-1, MMP-8 and MMP-9 polymorphisms, and levels of MMP-9 in preterm birth. Increased level of MMP-9 was reported in cases as compared to control. The significant association of MMP-9 (-1562) CT (P=0.001; OR = 1.44 (CI = 0.97–2.14)) and TT genotype (P=0.05; OR = 2.6 (CI = 1.46–4.69)) were reported in preterm birth. Our findings suggest that the MMP-9 plays an important role in contributing preterm labour and this can be used as a diagnostic tool during pregnancy. Keywords. infection; inflammation; matrix metalloproteinases; polymorphism; preterm birth. Introduction altered gene expression may be an attributing factor of causing preterm birth (Sheikh et al. 2016). The functional polymorphisms Preterm birth (PTB) is the most prevailing and persistent situated in MMP-1, MMP-8 and MMP-9 promoter region may be problem causing enormous morbidity and mortality among a contributing element (Fanjul-Ferna´ndez et al.
    [Show full text]
  • A Therapeutic Role for MMP Inhibitors in Lung Diseases?
    ERJ Express. Published on June 9, 2011 as doi: 10.1183/09031936.00027411 A therapeutic role for MMP inhibitors in lung diseases? Roosmarijn E. Vandenbroucke1,2, Eline Dejonckheere1,2 and Claude Libert1,2,* 1Department for Molecular Biomedical Research, VIB, Ghent, Belgium 2Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium *Corresponding author. Mailing address: DBMR, VIB & Ghent University Technologiepark 927 B-9052 Ghent (Zwijnaarde) Belgium Phone: +32-9-3313700 Fax: +32-9-3313609 E-mail: [email protected] 1 Copyright 2011 by the European Respiratory Society. A therapeutic role for MMP inhibitors in lung diseases? Abstract Disruption of the balance between matrix metalloproteinases and their endogenous inhibitors is considered a key event in the development of pulmonary diseases such as chronic obstructive pulmonary disease, asthma, interstitial lung diseases and lung cancer. This imbalance often results in elevated net MMP activity, making MMP inhibition an attractive therapeutic strategy. Although promising results have been obtained, the lack of selective MMP inhibitors together with the limited knowledge about the exact functions of a particular MMP hampers the clinical application. This review discusses the involvement of different MMPs in lung disorders and future opportunities and limitations of therapeutic MMP inhibition. 1. Introduction The family of matrix metalloproteinases (MMPs) is a protein family of zinc dependent endopeptidases. They can be classified into subgroups based on structure (Figure 1), subcellular location and/or function [1, 2]. Although it was originally believed that they are mainly involved in extracellular matrix (ECM) cleavage, MMPs have a much wider substrate repertoire, and their specific processing of bioactive molecules is their most important in vivo role [3, 4].
    [Show full text]