cell adhesion lecture eph 2014 láng, orsolya md, phd dept. genetics, cell & immunobiology,...
TRANSCRIPT
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Cell adhesion
www.dgci.sote.huLecture EPh 2014
Láng, Orsolya MD, PhDDept. Genetics, Cell & Immunobiology, Semmelweis University
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Cell adhesion
Contact with other cells and extracellular matrix
Role:
- embryonic development: formation of layers, tubes
- connection and communication between cells
- barrier, membrane polarity, mechanical attachment
- cell motility
- signal transduction
- cancer progression
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Cell-cell adhesion molecules1. Cadherins
2. CAMs or Ig like adhesion molecules
3. selectins
Cell-ECM adhesion Molecules4. Integrins
5. Proteoglycans (dystroglycan)
Homophyl or heterophyl
Ion dependency :Ca++, Mg++
Cytoskeletal component
Junctional complex
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Cadherins
Homophil connection Cell – cell
Ca++ - dependent binding
I.c. anchored to actin or intermedier filamentum
Significant role in developmentof vertebrates
Cell specificity
N – neuralP – placentalE - epithelial
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Cadherin superfamily
Type Localisation Adherent
cla
ssic
al
E Epithelial Adherens junction
N Neuronal, muscle, fibroblast
Adherens junction
Synapsism
P Placenta, epidermis, mammary gland
Adherens junction
VE Endothelial Adherens junction
Non
-cl
Desmocollin Skin Desmosomes
Desmoglein Skin Desmosomes
T cadherin Neuronal, muscle -
Cadherin 23 Inner ear Stereocilia
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Cadherins allow cells to sort themselves
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Ig-like adhesion molecules Homophil connection - typical Heterophil connection - rare
5 pcs. Ig-type domain Ca 2+ indep. adhesion
Linked to actin filaments
More than 20 variations Expressed in the critical phase of development
Tissue-specificity:N-CAM - neuronL-CAM – liverV-CAM1- vascular
In melanoma – relation between In melanoma – relation between ICAM-1 density and theICAM-1 density and themetastatic activity of the tumormetastatic activity of the tumorIntracellul
ar
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Lectin type proteins,Carbohydrate specificity
Tissue-specificity:E - epithelialL - lymphoidP - placenta
L selectin: it has role in the initial phase of adhesion;
in newborns the level of L sel. is low – the low number of inflammations (?!)
Selectin molecules
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Heterophil connection
Ca2+-dependent binding
Focal contacts Its i.c. linker proteins arei.e.. talin, -actinin, vinculin RGD sequ. is significant in ECM binding
Partner molecules: fibronectin
laminincollagen
Deficiency (Deficiency () – the adhesion of ) – the adhesion of leukocytes affected, resultsleukocytes affected, resultsthe increase of inflammations the increase of inflammations
Integrin molecules
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Type of integrins
Integrin Ligand Localisation Mutation of α or ß subunit
α5ß1 fibronectin ubiquitous Early death of the embryo,
α6ß1 laminin ubiquitous Early death of the embryo, severe skin blistering
α7ß1 laminin muscle Musclar dystrophy
αLß2 ICAM leukocytes Impaired recruitment of leukocytes, LAD, recurrent infections
αIIbß3 fibrinogen thrombocyte bleeding, no platelet aggregation, (osteoporosis)
α6ß4 laminin Hemidesmosomeepithelial cells
severe skin blistering
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Role of adhesion molecule in extravasation of leukocytes
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Adhesion molecules
www.cbrinstitute.org/.../media/image1.html
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Functional classification of cell junction
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Anchoring – mechanical junction
Barrier formation - separation
Channel like (gap junction) – communication
Signal relaying junctions sinapsm
immunological sinapsism
transmembrane proteins
Functional classification of cell junction
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Anchoring junctions
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Junctional complexes
Tight junction
Zonula adherens
Desmosome
Gap junction
Interdigitation
Hemidesmosome
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Isolate parts of plasma membrane (apical and basolateral) Completely encircle polarized cells Look like honey comb
Tight junction(TJ) = zonula occludens
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Claudins and occludins (membrane proteins) zip two membranes together
Molecular structure of tight junction
Stabilized by spectrinConnected to spectrin by adapter proteins ZO1 and ZO2
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Regulation of tight junctions
BeltsBelts of proteins that close extracellular space between cells Prevent passagePrevent passage of water and water-soluble substances Account for electrical resistance across epithelia
Leaky epithelia where there is need for some trafficHormonesVasopressinCytokinesLack of ATP causes “leak”Extravasating leukocytes open tight junctions
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Tight junctions separate components of the plasma membrane as well
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Mechanical juncion: zonula adherens - adhesion belt
Adherent junctionsHold cells tightly togetherConfer mechanical strengthCommon in tissue that are subject to severestress such as skin and cardiac muscle
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Molecular structure of zonula adherens
Belt like junctions located just below tight junction Simple points of attachment, do not contain channels connecting the interiors of the two attached cells
Adhesion molecule -cadherin (E)
Linker proteins - α, and catenin, vinculin
Cytoskeletal component - microfilament (actin)
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Desmosome = macula adherens
Cytoskeletal componentIntermedier filaments
(i.e. keratin)
Cytoplasmatic plaque
Adhesion moleculecadherins
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Components of desmosomes
The expression pattern of DSGs and DSC is tissue-specific and may even vary within one tissue, like in the different layers of the epidermis
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Spinous layer desmosome
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Focal adhesion
Hemidesmosome – Cell-ECM junction
Fixing of epithelial cells to the basal membrane
Adhesion molecule - integrins
/Linker proteins – plectin, dystonin/
Cytoskeletal component – intermediate filament (keratin)
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Desmosomes and hemidesmosomes both link to
intermediate filaments
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Gap junction – channel forming junction
Communicating junction
100-1000 connexon
protein: connexin
homomeric-heteromeric combinations
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Gap junction TEM SEM
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In plant cells plasmodesmata perform many of the same functions as gap junctions
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Interdigitation
Wave-like plasma membrane extensions + desmosomes
Characteristic of epithelial cell
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Basal striationnucleus
mitochondria
Basement
membrane
Basa
l st
riati
on
Increases the surface of the basal membraen for molecular transport
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ECM- Extracellular matrix
• Mechanical
• Influence on migration of the cells
• Regulation of activity of molecules released
• Co-receptors
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Main components of ECM
Protein componentGlycan
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Molecular Biology of the Cell (© Garland Science 2008)
LNs are cross-shaped proteins.All LN isoforms contain α, β1 and β2 chains that are connected by disulfide bonds (18 isoform of laminin- diverse in tissue)LN binds to membrane receptors (integrins) of the overlying cells.LN attaches cells to the basal lamina.
LN contains binding sites for other components of the basal lamina: type IV collagen, heparin,
Laminin (LN)
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Collagens
Provide strength of ECMmaintains form of tissue
Most abundant protein in vertebrates Found as bundles throughout ECM Structure:
Rigid triple helix of 3 intertwined polypeptide chains Unusual aa compositionn (hydroxilation) Collagen fiber consists of numerous fibrils (molecules,
polypeptides)
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Structure and synthesis of collagene ER/Golgi: Pro-a-chains are produced, hydroxylated and glycosylated at selected Lys and Pro residues. The lack of vitamin C prevents hydroxylation → impaired fibril formation (scurvy).Processed pro-peptides assemble into triple-helical pro-collagen. Golgi: Disulphide bonds form between the N- and C-termini of procollagen. After exocytosis, N- and C-termini are trimmed, allowing fibril assembly
Fibrils (diam.10-300 nm) More than 15 types I., II., III., V., XI. – formation of fibrils IV., VII. – network IX., XII. – association of fibrils
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Collagen Assembly
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Matrix types produced by vertebrate cells
COLLAGENE
I ChS, DS fibroblastII fibronectin ChS integrin chondrocyteIII HS, Hep. hepatocyte
epithel
IV laminin HS, Hep. laminin rec. epithel,endothel,regenerating hepatocyte
V fibronectin HS, Hep. integrin resting fibrobl.VI fibronectin HS integrin resting fibrobl.
Anchor Proteogly.Receptor Cells
Abbr.: ChS – chondroitin sulfate; DS – dermatan sulfate; HS – heparan sulfate; Hep - heparin
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Fibroblast surrounded by collagen fibrils
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Osteogenesis imperfecta – clinical manifestation
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Tissue elasticity
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Elastic fibers permit long-range deformability and passive recoil. Elastic modulus is ~0.1 MPa. This function is crucial for arteries, lung, skin and other dynamic connective tissues that undergo cycles of extension and recoil.
The major component of elastic fibers is the thread-like protein elastin Fibrillins provide an outer structure for amorphous, cross-linked elastin.
During ageing, elastin is degraded and becomes inflexible.
Elastin
Tropoelastine→ cross link→ functional elastin
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Fibronectin
Glycoproteins Fibronectin is a dimer of two identical 250 kDa subunits Alternative splicing of one gene produces ~20 human FNs RGD (Arg-Gly-Asp) component is recognised by integrines Mediate the connection between the ECM and the cell membrane. FN exists as a soluble form (plasma FN) and cellular FN.
Plasma FN is predominantly produced in the liver.Cellular FN is deposited into the ECM by a cell-mediated
process, FN binds a variety of other proteins like integrins, heparin, collagen, fibrin.
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Molecular Biology of the Cell (© Garland Science 2008)
TEM structure of basal lamina ~ basement membrane
In epithelial membrane – prevent cancer cell invasionKidney – serves as a filtration barrier
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Molecular Biology of the Cell (© Garland Science 2008)
Molecular structure of basal lamina
Transmembrane
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HA is a large, unbranched and negatively charged polymer of repeating (2-25K) disaccharides.
it is synthesized freely at the plasma membrane by hyaluronan synthases.
Free HA is found in the ECM of migrating cells.
HA binds to cell surface receptors (CD44) HA binds proteoglycans.
Structure of Hyaluronan
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Structure of aggrecan aggregate
In cartilage the key proteoglycan is aggrecan (MW: 2 x 108)
At 40 nm intervals aggrecan core proteins are attached (assisted by a linker protein) to a decasaccharide sequence in hyaluronan
Attached to the aggrecan core protein are multiple GAGs
The major GAGs in aggrecan are chondroitin sulphate and keratin sulphate
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Aggrecan
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Rheumatoid artritis
Synovial fluidSerum
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Molecular structure of the focal contactMolecular structure of the focal contactFimbrinFimbrinNexilinNexilin
TensinTensinαα-Actinin-ActininTalinTalin
PaxillinPaxillinCaveolinCaveolin
ZyxinZyxinPalladinPalladinVinexinVinexinPonsinPonsin
Integrin, Syndecan-4, Leukocyte common antigenIntegrin, Syndecan-4, Leukocyte common antigen
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Function of the adhesion molecules
Adhesion molecule
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Summary I.
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Molecular Biology of the Cell (© Garland Science 2008)
Summary II.