Transcript for:
ECM Overview and Components

Title: URL Source: blob://pdf/15e6b014-eafc-46e0-8afd-f7a3d90cf2d2 Markdown Content: EXTRACELLULAR MATRIX # (ECM) ## Ferbian Milas Siswanto, S.KH., M.Sc., Ph.D. ## Departemen Kimia dan Biokimia ## FKIK - UNIKA ATMA JAYA EXTRACELLULAR MATRIX ( ECM ) The extracellular matrix (ECM) is a non -cellular three -dimensional macromolecular network composed of an array of multidomain macromolecules organized in a cell/tissue -specific manner . Component : structural protein (collagen, elastin), specialized protein (fibrillin, fibronectin, laminin), proteoglycan (glycosaminoglycans) .FUNCTIONS OF ECM 1. Mechanical support for cell anchorage and cell migration, and maintenance of cell polarity . 2. Control of cell growth, ECM components can regulate cell proliferation by signaling through cellular receptors of the integrin family . 3. Maintenance of cell differentiation, morphogenesis, and homeostasis . 4. Scaffolding for tissue renewal . The integrity of the basement membrane or the stroma of the parenchymal cells is critical for the regeneration of the tissues .COMPONENTS OF ECM The ECM is a dynamic, constantly remodeling macromolecules complex synthesis, and secreted locally, which assembles into a network in the spaces surrounding cells . The ECM is component of three groups of macromolecules : 1. Fibrous structural proteins, such as the collagens and elastin that provide tensile strength & recoil . 2. Adhesive glycoproteins, such as fibronectin and laminin, that connect the matrix elements to one another and to cells 3. Water hydrated gels such as proteoglycans and hyaluronic acid that provide resilience and lubrication .COLLAGEN The collagens are a family of proteins that comprise about 30% of total protein mass in the body and are found to varying degrees in all tissues and organs. To date, more than 30 types of collagens have been identified that are broadly subclassified by their function, domain structure, and supramolecular organization. The collagen family can be generally divided into fibril -forming collagens (fibrillar) and nonfibrillar collagens. Collagen fibers are the most abundant structural components of the ECM. Type I collagen composition: [ 1(I)] 2 2(I) TYPES OF COLLAGEN (I) TYPES OF COLLAGEN (2) Tropocollagen Covalent crosslink Extension peptides 20 to 35 kDa procollagen aminoproteinase procollagen carboxyproteinase Lysyl oxidase (nonenzymatic) Glycation of collagen in the ECM increases the cross -linking of collagen. > The end -product of glycation reactions are termed AGEs (Advanced Glycation End Products) > Cross -linking can lead to the accumulation of various plasma proteins in the walls of blood vessels, in particular accumulation of LDL that can contribute to atherogenesis. > Uptake of AGEs by endothelial cells and macrophages can activate the transcription factor NF -kB, generating a variety of cytokines and pro - inflammatory molecules. i. Affected individuals produce only 50% of the normal amount of type I collagen, produced by the normal allele ii. Mechanisms a) Splicing in 1 gene b) Null mutation in 1 gene # OSTEOGENESIS IMPERFECTA NONFIBRILLAR COLLAGENS Nonfibrillar collagens are a heterogeneous group containing triple -helical segments of variable length, interrupted by one or more intervening nonhelical segments. Major type Type IV collagen = assembles into a flexible mesh -like network in basement membranes. Composition: 1(IV) [ 2(IV)] 2 Anomalies in type IV collagen in the kidney basement membrane result in glomerular diseases, including Goodpastures syndrome . This is a rare autoimmune disease caused by the production of antibodies that specifically bind to type IV collagen of basement membranes. Non -collagenous Proteins : ELASTIN Elastin is a connective tissue protein responsible for extensibility properties and elastic recoi l in tissues. Distribution: lung, large arterial blood vessels, elastic ligaments, skin, etc. In contrast to collagen, only one genetic type of elastin is known (ELN gene), although variants arise by alternative splicing (13 variant). Elastin is synthesized as a soluble monomer of ~70 kDa called tropoelastin . Some of the prolines of tropoelastin are hydroxylated to hydroxyproline by prolyl hydroxylase DISEASES RELATED TO ELASTIN > Emphysema , due to elastase ( a protease with specificity for small hydrophobic residues, such as alanine and valine) digests elastin in the alveolar wall. FIBRILLIN Microfibrils are fine fiber -like strands 10 to 12 nm in diameter which provide a scaffold for the deposition of elastin in the ECM. Fibrillins are large glycoproteins (about 350 kDa ) that are major structura l component of these fibers. They are secrete d (subsequent to a proteolytic cleavage) into the ECM by fibrob lasts and become incorporated into the insolub le microfibrils. Fibrillin -1 is the main fibri ll in present. Other proteins inclu ding microfibril -associated glycoproteins (MAGPs) , fibulins , and members of the ADAMTS family are also associated with microfibrils. Fibri ll in microfibrils are found in elastic fibers and also in elastin -free bundles in the eye, kidney, and tendons. FIBRONECTIN Fibronectin is a major glycoprotein of the ECM, also foun d in a soluble form in plasma. It consists of two identical subunits, each of about 230 kDa , joine d by two disu lfide bridges near their C-terminals. Humans have only one FN gene but are subject to considerab le alternative splicing (20 transcripts). Fibronectin contains three types of repeating motifs (I, II, and III), which are organized into functional domains (at least seven); functions of these domains inclu de binding fibronectin, fibrin, collagen, and cell surfaces. The binding to cell surface receptors of the integrin type is mediated by the sequence Arg -Gly -Asp (RGD). Many malignant cells are devoid of surface -bound FN, although they possess FN receptors. The binding of these receptors to tissue FN facilitates metastasis. LAMININ Laminins are a family of noncollagenous glycoproteins found in basement membranes. They are large (850 kDa ) heterotrimeric molecules composed of , , and chains. To date, five , four , and three chains have been identified, which can associate to produce at least 15 different laminin variants. The three interacting chains in a heterotrimer are arranged in an asymmetric cruciform, or cross -shaped, molecule, held together by disulfide linkages. Like fibronectin, laminins interact with cells through multiple binding sites in several domains of the molecule. Laminin polymers are also connected to type IV collagen by a single chain protein ( nidogen/entactin ), which has a binding site for collagen, the heparan sulfate proteoglycan ( perlecan ), and also has an RGD sequence for integrin binding. Nidogen plays a central role in the formation of crosslinks between laminin and type IV collagen, generating a scaffold for anchoring of cells and ECM molecules in the basement membrane. Structure of the basal lamina . Laminin is attached to type IV collagen via nidogen and perlecan (forming the basal lamina) and to the epithelial cell layer via integrins and dystroglycan .PROTEOGLYCAN Proteoglycans are the gel -forming components of the ECM composed of core proteins containing covalently bound sugars (glycosaminoglycans [GAGs]) and these units form large complexes with other components of the ECM, such as hyaluronic acid or collagen. STRUCTURE OF GLYCOSAMINOGLYCANS GAGs are linear, unbranched oligosaccharides that may contain more than 100 sugar residues in a linear chain. GAGs have a disaccharide repeat which is different for each type of GAG but is usually composed of a hexosamine and a uronic acid residue, except in the case of keratan sulfate, in which the uronic acid is replaced by galactose. The amino sugar in GAGs is either glucosamine (GlcNH2) or galactosamine (GalNH2), both of which are present mostly in their N- acetylated forms ( GlcNAc and GalNAc ). In some of the GAGs (e.g., heparin, heparan sulfate), the amino group is sulfated rather than acetylated. The uronic acid is usually D-glucuronic acid ( GlcUA ), but in some cases (e.g., dermatan sulfate, heparin), it may be L-iduronic acid ( IdUA ). With the exception of hyaluronic acid and keratan sulfate, all the GAGs are attached to protein by a core trisaccharide, Gal -Gal -Xyl ; the xylose is linked to a serine or threonine residue of the core protein. Keratan sulfate is also attached to protein, but in that case, the linkage is through either an N-linked oligosaccharide (keratan sulfate I) or an O-linked oligosaccharide (keratan sulfate II). Hyaluronic acid, which has the longest polysaccharide chains, is the only GAG that is not attached to a core protein. FUNCTIONS OF THE PROTEOGLYCANS One of the major roles of proteoglycans is to provide structural support to tissues, especially cartilage and connective tissue. In cartilage, large aggregates, composed of chondroitin sulfate and keratan sulfate chains linked to their core proteins, are noncovalently associated with hyaluronic acid via link proteins, forming a jelly - like matrix in which the collagen fibers are embedded. This macromolecule, a bottlebrush structure known as aggrecan, provides both rigidity and stability to connective tissue SYNTHESIS OF PROTEOGLYCANS DEGRADATION OF ## PROTEOGLYCANS Lysosome s Protein portion lysosomal proteases GAG chains lysosomal hydrolases THANK YOU Ferbian Milas Siswanto, S.KH., M.Sc., Ph.D. Departemen Biokimia , FKIK -Unika Atma Jaya Email: [email protected] WA: 081337321736