E-mail: cgrasho@biochem.mpg.de
The three filament networks
intermediate
f-actin tubulin filaments
3
Many disorders are ‘mechanical diseases’ !
4
Cancer cells are mechanosensitive
5
Many biological processes are mechanosensitive
6
What we would like to understand
7
What we would like to understand
8
What characterizes the mechanical environment ?
- the mechanical
properties of the
cellular environment
are characterized by
the extracellular
matrix (ECM)
9
The extracellular matrix (ECM)
We distinguish:
- filamentous proteins
- glykosaminoglycanes
(GAGs)
- proteoglycanes
- adhesion proteins
10
Collagen: the most abundant EMC protein
- we know 28 different
collagen subtypes
consisting of 46 distinct
polypeptides
- collagens are
evolutionary conserved
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Collagen and the collagen triple helix
- collagen is characterized
by a typical amino acid
repeat:
(G-X-Y)n
G: glycine
X: proline
Y: hydroxy-proline
12
Collagen and the collagen triple helix
- collagen is characterized
by a typical amino acid
repeat:
(G-X-Y)n
G: glycine
X: proline
Y: hydroxy-proline
13
The role of hydroxyproline
(G-X-Y)n
prolin
X: often Prolin
Y: often Hydroxy-Prolin
- hydroxyprolin stabilizes
the triple-helix through
prolylhydroxylase
stereo-electronic effects
+ Vitamin C
- hydroxyproline is not a
natural amino acid and has
hydroxy- to be synthesized by the cell
prolin
- lack of hydroxyproline
leads to scurvy
14
Collagen fibers are very long
15
Collagen fibrillogenesis is a multi-step process
- lysyloxidase (LOX)
catalyzes the oxidation of
lysine groups
- LOX-expression correlates
with malignancy of breast
cancer
18
Defects in the collagen protein cause diseases
Scurvy Osteogenesis
(general) Imperfecta (Col1A)
The problem
20
Our tissue needs to be elastic !
21
Elastin gives tissues their elasticity
(P-G-V-G-V-A)n
23
Elastomeres and the elastic module
elastic modulus:
non-elastic
break
E=σ/ε
stress σ [ N / m2 ]
[ N / m2 ] = Pa
[ N / mm2 ] = MPa
[ kN / mm2 ] = GPa
The problem
27
What mediates compressibility ?
28
GAGs mediate compressibility
- glycosaminoglykane
(GAGs) consist of
repeating sugar subunits
hyaluronan
chrondoitin sulfate
dermatan sulfate
keratan sulfate
heparan sulfate
29
GAGs occupy a large volume
30
GAG example - Hyaluronan
- proteoglycanes are
composed of GAGs covalently
linked to a core protein
32
Proteoglycane - Aggrecan
33
Adhesive glycoproteins
sugars
elastomers
triple helix
37
What we would like to understand
38
Direct and indirect sensing mechanisms
- we distinguish:
cell-cell adhesions
cell-ECM adhesions
39
Overview of cell adhesion structures
40
Integrins mediate cell-matrix adhesions
41
Human cells express many integrins
a1 β extracellular
a2
a3 α
a4
a5
b1
a6
b2
a7
b3
a8
a9 b4
a10 b5
a11 b6
aV b7
aIIb b8
aD
aX
intracellular
aL
aM
aE
42
Integrin receptor classes
43
Integrins are very important
- without integrins
embryos die even
before implantation
into the ueterus !
44
Integrins are important for platelet activation
45
Integrins can be active or inactive
EZM
inactive
conformation active
conformation
Kindlin
The problem
48
Focal adhesions form at integrin tails
49
FAs affect many cellular processes
adapter proteins
kinases, phosphatases,
proteases, etc. cell division
cell migration
mechanical integration 50
Summary – cell adhesion and integrins
51
What we would like to understand
52
Integrin-dependent force transduction
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Talin can interact with integrins and actin
FERM domain
(integrin binding)
vinculin binding domain
dimerization domain
vinculin binding
domains
54
A model of force transduction in FAs
f-actin-talin
binding
55
A model of force transduction in FAs
56
A model of force transduction in FAs
vinculin
recruitment
57
A model of force transduction in FAs
vinculin
recruitment
to talin
58
A model of force transduction in FAs
59
A model of force transduction in FAs
integrin clustering
60
Measuring force transduction
The problem
61