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  • Cell Adhesion and Signaling

    黃敏銓 [email protected]

    Institute of Anatomy and Cell Biology 1

  • 2

    Transactivation

    NATURE REVIEWS | CANCER VOLUME 7 | FEBRUARY 2007 | 85

  • Functions of Cell Adhesion

    proliferation

    differentiation

    inflammation

    angiogenesis

    apoptosis

    polarity

    cell shape

    survival

    migration

    cell cycle

    Cell Adhesion

    3

  • 4

    CELL-CELL ADHESION Cadherins E, N, P

    cadherins Ca2+- or Mg2+ -dependence

    homophilic actin filaments (via catenins)

    adhesion belts

    desmosomal cadherins

    yes homophilic intermediate filaments (via desmoplakins, plakoglobin and other proteins)

    desmosomes

    Ig family members N-CAM, L1 no homophilic or heterophilic

    unknown no

    Selectins (blood cells + endothelial cells)

    P-selectin yes heterophilic unknown no

    Integrins (on blood cells) LFA-1 (aLb2), Mac-1 (aMb2)

    yes heterophilic actin filaments no

    CELL-MATRIX ADHESION Integrins many types yes heterophilic actin filaments

    (via talin, vinculin, and other proteins)

    focal contacts

    a6b4 yes heterophilic intermediate filaments

    hemidesmosomes

    Transmembrane Proteoglycans

    syndecans no heterophilic actin filaments no

  • Major families of cell-adhesion molecules

    5

  • 6

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

  • 7

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

  • Significance of cytoskeletons in cell adhesion

    8

    No effect on cell

    Cell adhesion

    without cytoskeletal attachment

    with cytoskeletal attachment

    PULL

    PULL

    cytoskeleton

    adhesion molecule Plasma membrane

  • 9

    1. Tethering and Rolling

    3. Firm adhesion

    4. Transmigration

    Inflammatory stimulus

    2. Activation

    Selectins

    Chemoattractants

    Integrins

    endothelium

    Ig CAMs

    9

  • Selectin

    10

    Short consensus repeat

    EGF-like domain

    Lectin domain

    L-selectin

    E-selectin

    P-selectin

  • 11

    L-Selectin

    LEUKOCYTE

    P-Selectin E-Selectin

    ENDOTHELIAL CELLS

    Thrombin Histamine TNF- α LPS

    TNF- α IL-1 β IFN- γ LPS

  • ESL-1 PSGL-1L-Selectin

    LEUKOCYTE

    h

    ENDOTHELIAL CELL

    GlyCAM-1 CD34 MAdCAM-1 E-Selectin P-Selectin

    12

  • Protein Carbohydrate

    INTERACTION

    Selectin Selectin Ligand

    on lipids

    proteins

    13

  • Summary of intracellular signaling cascades in leukocytes on L-selectin triggering

    p56Lck

    cell growth, survival, apoptosis

    P-Ser

    (J. Biol. Chem., V. 279, 34472- 34480, August 13, 2004)

    PKC

    14

    Adapted from Pediatric Research 48:283-288 (2000)

  • Molecular Events in Transmembrane Signaling via E-selectin

    E-selectin

    SHP2 Shc

    Grb2

    Sos

    Ras Raf MEK

    ERK1/2

    (HUVEC cell) -Tyr-PP-Tyr603-

    15 gene expression

    (ex: c-fos) J. Biol. Chem., Vol. 276, Issue 51, 48549-48553, December 21, 2001

  • Regulation of transendothelial migration of colon cancer cells by E-selectin-mediated activation of p38 and ERK MAP kinases

    Tremblay PL Auger FA

    16

    , , Huot J. Oncogene. 2006 Oct 26;25(50):6563-73.

    http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_AbstractPlus&term=%22Tremblay+PL%22%5BAuthor%5D http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_AbstractPlus&term=%22Auger+FA%22%5BAuthor%5D http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_AbstractPlus&term=%22Huot+J%22%5BAuthor%5D

  • Signaling cascades initiated by ligation of either the selectins or their ligands

    Calcium flux Src family kinases Ras activation MAP Kinases Neutral sphingomyelinase

    Gene expression (ex: TNFα, IL-8, CXCR4)

    Oxygen radical generationCell shape change

    β2 integrin activation

    17

    Adapted from seminars in IMMUNOLOGY, Vol. 14, 2002: pp. 73–81

  • Integrin

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

  • Signaling transduction pathways emanating from Integrins

    31 MAY 2002 VOL 296 p.1652-1653 SCIENCE

    19

  • 20

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

  • Common integrins and ECM ligands bound

    Journal of Cell Science 115, 3729-3738 © 2002

    21

  • Integrin binding proteins

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

    22

  • Integrin binding proteins

    23

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

  • Integrin Signaling Pathway

    Integrins

    24

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

    http://stke.sciencemag.org/

  • FAK Focal Adhesion Kinase

    25

  • 26

    Oncogene (2004) 23, 7928–7946.

  • Focal Adhesion Kinase (FAK) nonreceptor tyrosine kinase

    27

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

  • 28

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

    Example of FAK function

    Focal Adhesion Kinase

    ERK ERK

    JNK

    CrkII

    C3G

    Rap-1

    B-Raf

    Raf

    Shc

    survival

    contractibility

    P P

    caveolin

    Ras Ras

  • Multiple signaling proteins dock at Tyrosine 397 of FAK

    Biochimica et Biophysica Acta 1540 (2001) 1-21

    pY397

    29

  • 30

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

  • Scanning electron micrograph of a basal lamina in the cornea of a chick embryo

    Some of the epithelial cells (E) have been removed to expose the upper surface of the matlike basal lamina (BL). A network of collagen fibrils (C) in the underlying connective tissue interacts with the lower face of the lamina. (Courtesy of Robert Trelstad.) 31

  • a c Mock β4GalNAc-T3

    Mock β4GalNAc-T3

    b d

    32

  • a Mock β4GalNAc-T3

    0 h

    32 h

    Mock β4GalNAc-T3 0 2 4 6 8

    10 *

    M ig

    ra tio

    n ra

    te (μ

    m /h

    )

    33

  • b Mock β4GalNAc-T3

    Mock β4GalNAc-T3

    34

  • B Mock

    IP: FAK (C20)

    4G10 (anti-pY)

    FAK

    IB:β4GalNAc-T3 A

    1.0 2.7

    β4GalNAc-T3 IB:Mock β4GalNAc-T3

    1.0 2.5 1.0 2.5

    1.0 1.7

    pY397

    pY576

    35

    IP: FAK (C20)

    FAK

    pY118-paxillin

    IB:

    paxillin

    pY31-paxillin

    Mock β4GalNAc-T3 C

    1.0 1.8

    1.0 0.5

    PY20 (anti-pY)

    1.0 3.5

    D β4GalNAc-T3Mock

    1.0 1.8

    pERK1/2

    ERK1/2

    β-actin

    WCL

  • ILK Integrin-Linked Kinase

    36

  • Integrin-Linked Kinase binding proteins

    37

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

  • 38

    ILK-PINCH-parvin (IPP) complex

    NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 7 | JANUARY 2006 | 22

  • Signaling through IPP complex

    39

    NATURE REVIEWS | MOLECULAR CELL BIOLOGY VOLUME 7 | JANUARY 2006 | 24

  • NATURE REVIEWS | CANCER VOLUME 5 | JANUARY 2005 | 60 40

  • Activation of MAP kinase cascades by integrins

    41

  • Activation of MAP kinase cascades by integrins

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

    Focal Adhesion Kinase

    ERK ERK

    JNK

    CrkII

    C3G

    Rap-1

    B-Raf

    Raf

    Shc

    survival

    contractibility

    P P

    caveolin

    Ras Ras

    42

  • Integrin regulates actin filaments formation via Rho GTPases

    (Rho, Rac, CDC42)

    43

  • Integrin signaling to the cytoskeleton via Rho GTPases

    44

    Rho Rac CDC42

    cofilin P

    STRESS FIBERS

    Lamell ipodia

    FIL OP

    OD IA

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

  • Mechanism of activation of N-WASP

    P Active N-WASP

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

  • Mechanism of activation of WAVE2

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

  • Cooperativity between growth-factor receptors and integrins

    NATURE CELL BIOLOGY VOL 2 DECEMBER 2000 E225-229

    growth factor receptorintegrin

    47

  • 48

    1

    2

    3

    integrin Receptor Tyrosine Kinase

    Integrin regulates signaling in RTK/Ras/MAPK pathway

    cytoplasm