Cell adhesion and ECM
description
Transcript of Cell adhesion and ECM
![Page 1: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/1.jpg)
Lecture 10:
Cell Biology interactive media ”video” or ”animation”
The eukaryotic kingdom
1
Cell biology 2014 (revised 12/2-13)
![Page 2: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/2.jpg)
The four major tissues in the human body
- Epithelial
- Muscle
- Connective
- Nerve
Cells
Cells + ECM
Metazoan cells form organs with specialized tissues:
2
![Page 3: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/3.jpg)
Different types of cell adhesion
Homophilic binding Heterophilic binding
3
![Page 4: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/4.jpg)
Cell-cell contacts in columnar epithelia
Gap junction
Tight junction
Adherens junction
Desmosome
Connection allowing local communication
Cell-cell adhesion
4
hemidesmosome Cell-ECM adhesion
Basal lamina
Restricting movement of extra-cellular fluids
![Page 5: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/5.jpg)
Tight junctions seal epithelial sheets to block passage of fluids in between cells
I. Tight junctions: function
Intestine
Blood vessels
Glucose
GlucoseActive and selective transport through the cytosol of cells by e.g., the Na+ driven glucose symport
5
![Page 6: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/6.jpg)
II. Tight junctions: Architecture
The appearance of tight junctionsresemble stitches across the plasma membranes of the two cells
Tight junctions are made up by occludin and claudin. These are transmembrane proteins, which form tight connections across the extracellular space
Linking protein attaches occludin and claudin to the cortical actin cytoskeleton
6
![Page 7: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/7.jpg)
P.M.
a-catenin
a-catenin
Cadherin (calcium-dependent adhesion)
Cell #1
Cell #2
P.M.
b-catenin
b-catenin
Actinfilament
Many cadherins are known: E-cadherin in EpitheliaN-cadherin in Neural cells
Linkers of cadherinsto the actin cytoskeleton
Adherence junctions from stable cell-cell adhesion points between adjacent cells
video 19.1- adhesion_junctions7
Ca2+
I. Cadherins: adherence junctions
![Page 8: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/8.jpg)
II. Cadherins: growth arrest at cell-cell contact
P.M.
G1
Wnt
G1 cyclin gene
..but is stabilized by Wnt signaling
a-cateninb-catenin
b-catenin
TCF
b-catenin
Cytosolic b-catenin is by default unstable.....(Ubiq. dep. degradation)
1.
2.
3. b-catenin enters the nucleus: G1 cyclin transcription cell proliferation
4. Sequestering of cytosolic b-catenin at the adherence junctions formed after cell proliferation (i.e., at ”density arrest”)
8
Ca2+
![Page 9: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/9.jpg)
III. Cadherins: organization of cells into organs
Cells expressing different cadherins
Cells expressing different amounts ofthe same cadherin
Cadherins are important for organ formation during development9
+Ca2+
+Ca2+
![Page 10: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/10.jpg)
Desmosomes hold cells togetherlike rivets. Through linkage toIFs, they distribute shear forcesevenly within the cell
P.M.
Cell #1
Cell #2
P.M.
Linkers
Linkers
Intermediatefilament (IF)
Cadherin family protein
Structure and function of the desmosome
animation 16.4- intermediate_filament 10
Ca2+
![Page 11: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/11.jpg)
Structure and regulation of gap junctionsConnexon
= ~1.5 nmFree passage of: Amino acids Nucleotides Sugars Ions ”2nd messengers”
Connexin
P PP PP P
Different connexins –different pore size
Cell #1
Cell #2Regulation of pore size
11
![Page 12: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/12.jpg)
I. Integrins: Structure and ligand specificityi) Hetero-dimeric proteins consisting of a- and b- chains ii) At least 21 cell-type specific isoforms of a/b-chain pairsiii) Integrin ligands include ECM components (collagen, fibronectin, laminin) and structures on neighboring cells
12
ECM: Basal laminaECM: connective tissue
bx ax (ligand: laminin)by ay (ligand: fibronectin)
Integrins linked to IF (hemidesmosomes: epithelia)
Integrins linked to actin (focal adhesions: fibroblasts)
![Page 13: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/13.jpg)
II. Integrins: Anchorage to ECM
ECM: connective tissue (contains residual migratory cells)
13
ECM: Basal lamina
Inactiveintegrin
Integrins linked to IF (hemidesmosomes) Static cell-ECM interactions , e.g. epithelial sheets
Integrins linked to actin (focal adhesions) Dynamic cell-ECM interactions, e.g., during migration of fibroblasts or leukocytes
Basal lamina: barrier towards connective tissue
![Page 14: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/14.jpg)
III. Integrins: Architecture of the focal adhesion
ECM
P. M.
Active integrin
Focal adhesions exist only in motile cells (i.e., not in epithelia) The dynamic nature of focal adhesion is dependent on both
“Inside-out” and “Outside-in” signaling
FAK: Focal adhesion kinase integrin dependant signaling
14
recruitment of SH2-domain signaling proteins (Clustering of FAK trans-phosphorylation, i.e. the same principle as for tyrosine kinase receptors, which are dimerized by ligand binding)
TalinLinkerFAK
Tyr- P
TalinFAK
Tyr- P
Tyr- P
![Page 15: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/15.jpg)
IV. Integrins: Regulation of ligand-affinity
15
Outside-in activation of ECM-binding
Inside-out activation of ECM-binding
1. Default state: The a- and b-chains are tightly associated Activated state: a- and b-chains are pushedapart and clustered by talin High affinity/avidity ECM-association
1.
2.
2.
The a- and b-chains of integrins have affinity for both “each other” and ECM ligands the concept of competing affinities
![Page 16: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/16.jpg)
V. Integrins: Inside-out activation
FAK
P
2.
1.
3.
2.
3.
Activated talin:i) Pushes a- and b-chains apartii) Clusters the cytosolic parts of integrin b-chains iii) Links b-chains with actin filamentsiv) Recruit focal adhesions proteins (vinculin, FAK etc) generation of a focal adhesion point
2.
1.
3.
4.
Activation of talin by a RTK ligand (e.g. EGF)
Separation of a- and b-chains
High affinity ECM-binding
Integrin clustering increased avidity
Albert et al. Fig 19-49
4.
16
Inactive talin
![Page 17: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/17.jpg)
VI. Integrins: Outside-in activationBinding by (very) high affinity ECM ligands……..breaks the interaction between the a- and b-chainsThe exposed b-chain talin-binding site……..….activates talin Generation of a focal adhesion point
outside-in + inside-out = positive feedback
1.
2.
3.
4.
ECM
1.
ECM
2.
2.
3.
inactivetalin
ECM
4.
17
![Page 18: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/18.jpg)
VII. Integrins: survival and cell proliferation signals
Plasma membrane
MAPK P
P
PPKB/Akt
P
PI-3 K
P
myc
Cell cycle entry
FAK
BadP
14-3-3
18
RasGTP
Motile cell types requires ECM for both growth and survival
Survival
PP3
-Tyr
- P
-Tyr
- P-T
yr- P
![Page 19: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/19.jpg)
- Provides mechanical support to tissues
- Organizes cells into tissues
- ‘Instructs’ cells as to where they are and what they should do
- Reservoir for extra-cellular signaling molecules
I. The extra-cellular matrix (ECM)
19
![Page 20: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/20.jpg)
II. The extracellular matrix (ECM)
Proteoglycan molecules form highly hydrated gel-like “ground substance” in which the fibrous proteins are embedded
Structural proteins, such as collagen and elastin, strengthen and organize the matrix
Composed of polymeric networksof several types of macromolecules. Secreted by connective tissue cells,such as fibroblasts & chondrocytes.
1. 2.
3.
1.
2.
3. Multi-adhesive proteins, such as fibronectin and laminin, facilitate cell attachment to the ECM
The aqueous phase of the ECM permits diffusion of nutrients20
![Page 21: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/21.jpg)
III. ECM: general structure of proteoglycan
H2O
Na+
--
--
-
Ca2+
Protein core
Polysaccharide sidechain
Negatively charged saccharides attract counter ionsand water, giving the ECM the property to resist compression and bounce back to its original shape
Osmosis
Linking saccharidesGlucosamino-glycans (GAGs)linear polymers of repeating disaccharides
O-linked sugar
21
![Page 22: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/22.jpg)
IV. ECM: Proteoglycan aggregates
Hyaluronan, up to 50 000repeating disaccharides
Linker protein
Proteoglycans can form huge aggregates onto hyaluronan. These aggregates can be up to 4 mm in length
These aggregates have a very high shock absorbing capacity and are highly enriched in cartilage 22
![Page 23: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/23.jpg)
V. ECM: Collagen architecture
Collagen a-chain (single helix)
Collagen molecule(triple helix)
Collagen fibril
Collagen fiber
Assembled outside the cell
Assembled in ER
Collagen is the most common protein in body, it forms strong andflexible fibers. Many types (at least 15)
23
![Page 24: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/24.jpg)
VI. ECM: Elastic elastin networks
Single elastin molecule
In cases there ECM is very flexible, e.g., in skin, lungs and blood vessel walls, some of the collagen is replaced by elastin.Cross-linked elastin behaves like a rubber band!
Stretching Relaxation
Crosslinking
24
![Page 25: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/25.jpg)
VII. ECM: different types of connective tissue
”Normal” connective tissue Cartilage
Fibroblast Chondrocyte
Ca10(PO4)10(OH)2
Ca10(PO4)10(OH)2
Bone
Osteoblast
Physical properties of the tissue depend on the content of the ECM, which is determined by the residual cell type 25
![Page 26: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/26.jpg)
Summary: ECM – a sticky business!
26
Laminin - Present in basal lamina of epithelia and the ligand for hemidesmosomes
Fibronectin - Present in all ECM and primary high-affinity ligand for focal adhesions
![Page 27: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/27.jpg)
Fig. 19-1: Epithelial tissue: The intermediate filaments of the cells themselves (linked from cell to cell by desmosomes) provides mechanical strength. Hemidesmosomes (integrin binding to laminin) are only found in the epithelial cells that connect to the basal lamina. These epithelial cells are normally essentially non-motile.
Connective tissue: ECM provides the mechanical strength, the sole role of the residual cells (fibroblasts) is to produce the ECM components. These residual cells move around and may migrate to e.g. a site of tissue damage. 27
Differential means to achieve mechanical strength
Epithelial cells
Basal lamina(dense ECM)
Connective tissue(ECM + cells)
Cells resistant to mechanical stress
ECM (but not cells) resistent to mechanical stress
![Page 28: Cell adhesion and ECM](https://reader035.fdocuments.us/reader035/viewer/2022081517/568163bb550346895dd4d27b/html5/thumbnails/28.jpg)
“Recommended reading”
Alberts et al5th edition
Chapter 191131-11451150-11621164-1194
Focus on the general principlesand topics highlighted inthe lecture synopsis
28