Cell AdhesionCell Adhesion

黃敏銓黃敏銓mchuang@ntu.edu.tw

G d t I tit t f A t d C ll Bi l1

Graduate Institute of Anatomy and Cell Biology

CellCell--Cell Adhesion and CellCell Adhesion and Cell--Matrix AdhesionMatrix Adhesion

actinfilaments

cadherinadhesion belt(cadherins)

filaments

Ig CAMs

integrinl tiselectin

focal contactintegrin

focal contact(integrins)

2

Functions of Cell Adhesioncell cycle

proliferation migration

differentiation survival

Cell Adhesion

inflammation cell shape

angiogenesis polarity

3apoptosis

CELL-CELL ADHESIONCadherins E, N, P Ca2+- or Mg2+

-dependencehomophilic actin filaments (via adhesion belts

cadherins -dependence catenins)

desmosomal cadherins

yes homophilic intermediate filaments (via desmoplakins,

desmosomes

desmoplakins, plakoglobin and other proteins)

Ig family members N-CAM, L1 no homophilic or h t hili

unknown noheterophilic

Selectins (blood cells + endothelial cells)

P-selectin yes heterophilic unknown no

Integrins (on blood cells) LFA 1 (a b ) yes heterophilic actin filaments noIntegrins (on blood cells) LFA-1 (aLb2), Mac-1 (aMb2)

yes heterophilic actin filaments no

CELL-MATRIX ADHESIONIntegrins many types yes heterophilic actin filaments

(via talin, vinculin, and other proteins)

focal contacts

proteins)

a6b4 yes heterophilic intermediate filaments

hemidesmosomes

Transmembrane syndecans no heterophilic actin filaments no

4

Transmembrane Proteoglycans

syndecans no heterophilic actin filaments no

Major families of cell-adhesion molecules

5

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Significance of cytoskeletons in cell adhesion

ith tadhesion molecule

Plasma membrane

No effect on cellwithout

cytoskeletalattachment

PULL

cytoskeleton

withcytoskeletal

Cell adhesiony

attachment

8PULL

Cell-Cell adhesionCell Cell adhesion

9

Ig CAMs

endothelium

109

SelectinP-selectin

EGF-like domain

Lectin domain

E-selectinse ec

Short consensus repeatL-selectin

Short consensus repeat

11

LEUKOCYTE

P Selectin

L-Selectin

P-Selectin E-Selectin

ENDOTHELIAL CELLS

ThrombinHistamineTNF- LPS

TNF- IL-1 IFN-

12

LPS LPS

L-Selectin ESL-1 PSGL-1

LEUKOCYTE

L-Selectin ESL 1 PSGL 1

h

ENDOTHELIAL CELL

E-SelectinGlyCAM-1 MAdCAM-1CD34 P-Selectin

13

INTERACTIONINTERACTION

Selectin Selectin Ligandlipids

Protein Carbohydrate on lipids

proteinsp

14

Leukocyte Adhesion Deficiency II (LAD II)

I t t i th t d tiIntrauterine growth retardation

Recurrent infection

Mental retardation

Short statureNat Genet. 2001. 28:69-72

Mutation in GDP-fucose transporterAutistic features

Convulsions

Mutation in GDP fucose transportercauses no fucose at the cell surface.

Cerebral atrophy

Microcephaly

a 20-year-old manwith LAD II

Coarse face

J Clin Immunol. 2010 Mar;30(2):308-13.15

Leukocyte Adhesion Deficiency IIIy y

16

Nature Medicine 15, 249 - 250 (2009)

Cell-ECM adhesionCell-ECM adhesion

17

IntegrinIntegrin

18Cell, Vol. 110, 673–687, September 20, 2002

Signaling transduction pathways emanating from Integrinsfrom Integrins

1931 MAY 2002 VOL 296 p.1652-1653 SCIENCE

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NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 2 | NOVEMBER 2001 | 795

Common integrins and ECM ligands bound

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Journal of Cell Science 115, 3729-3738 © 2002

Integrin binding proteins

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Integrin binding proteinsIntegrin binding proteins

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Integrin Signaling Pathway

Integrins

24

http://stke.sciencemag.org May 31, 2002. J. Thomas Parsons

FAKFAKFocal Adhesion Kinase

25

26

Oncogene (2004) 23, 7928–7946.

Focal Adhesion Kinase (FAK)t t i kinonreceptor tyrosine kinase

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NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 6 | JANUARY 2005 | 59

Example of FAK function

caveolin

Focal Adhesion Kinase ShcP P

CrkII

C3GRas

Ras

Rap-1 Raf

survival

ERK

B-Raf

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Modified from Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

ERKERK

contractibility

Multiple signaling proteins dock at Tyrosine 397 of FAKTyrosine 397 of FAK

pY397p

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Biochimica et Biophysica Acta 1540 (2001) 1-21

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NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 6 | JANUARY 2005 | 58

Mock IB:β4GalNAc T3A B

Mock

4G10

IB:β4GalNAc-T3

pY397

IB:Mock β4GalNAc-T3

1.0 2.5

β4GalNAc-T3

1.0 2.54G10(anti-pY)

1.0 2.7

FAK

pY576

1.0 1.7PY20(anti pY)

FAKIP: FAK (C20)

FAK(anti-pY)1.0 3.5

DIP: FAK (C20)

IB:Mock β4GalNAc-T3C

D

pERK1/2

Mock β4GalNAc-T3

pY118-paxillin

pY31-paxillin

1.0 1.8 ERK1/2

1.0 1.8

paxillin

pY31 paxillin

1.0 0.5

WCL

β-actin

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ILKILKIntegrin-Linked Kinase

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Integrin Linked Kinase binding proteinsIntegrin-Linked Kinase binding proteins

33

The Journal of Cell Biology, Volume 155, Number 4, November 12, 2001 505-510

ILK-PINCH-parvin (IPP) complex

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Signaling through IPP complex

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36NATURE REVIEWS | CANCER VOLUME 5 | JANUARY 2005 | 60

Activation of MAP kinaseActivation of MAP kinase cascades by integrinscascades by integrins

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Activation of MAP kinase cascades by integrins

caveolin

Focal Adhesion Kinase ShcP P

CrkII

C3GRas

Ras C3G

Rap-1 Raf

survival

Ras

ERK

B-Raf

survival

38

Modified from Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

ERKERK

contractibility

Integrin regulates actin filamentsIntegrin regulates actin filamentsformation via Rho GTPases

(Rh R CDC42)(Rho, Rac, CDC42)

39

Integrin signaling to the cytoskeleton via Rho GTPases

Rho Rac CDC42

Pcofilin

STRESS FIBERS

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

Mechanism of activation of N-WASP

P Active N-WASP

41Adapted from Journal of Cell Science 114, 1801-1809, 2001

Mechanism of activation of WAVE2

42Journal of Cell Science 114, 1801-1809, 2001

Cooperativity between growth-factor receptors and integrins

growth factor receptorintegrin

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NATURE CELL BIOLOGY VOL 2 DECEMBER 2000 E225-229

Integrin regulates signaling in RTK/Ras/MAPK pathway

integrin Receptor Tyrosine Kinase

1

2

cytoplasm1 cytoplasm

33

nucleus

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323

1. Activation of receptor tyrosine kinase (RTK)

I t i ti d li d bi di• Integrin aggregation and ligand binding

– EGFR, PDGF-R, FGFR phosphorylation

• Integrin aggregation alone

– PDGF-R, EGFR

• Direct integrin-RTK interaction

– v3 and PDGF-R, insulin receptor,

VEGF-R2

• Via integrin-associated cytoskeletal

components

– Integrin-actin-Neu RTK, integrin-FAK-

45EGFR or PDGFR

Integrin regulation of growth factor receptors

46

NATURE CELL BIOLOGY VOL4 APRIL 2002 E75-76

FAK integrates growth-factor and integrin signals to promote cell migrationg p g

47Current Opinion in Genetics & Development 2004, 14:92–101

2. Coupling between upstream and downstream molecules

Between Ras-Raf

– EGFR, PDGF-R in

anchorage-dependent

cell growthcell growth

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3. Transmission of the signal from cytoplasm to nucleus

J Cell Biol 2001 Apr 16;153(2):273-82

• Integrin mediated adhesion• Integrin-mediated adhesion Regulates ERK Nuclear Translocation andTranslocation and Phosphorylation of Elk-1– Actin cytoskeleton-

dependent (NIH 3T3 cells)

– G1 phase progression

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Examples of synergisms between integrins and RTK i liRTK signaling

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Annu. Rev. Pharmacol. Toxicol. 2002. 42:283–323