Junctions Flashcards
Integrins
- Transmembrane proteins that tie the ECM to cell cytoskeleton
- Integrins bind their ligands with low affinity, but are present in high numbers on the cell surface.
Integrin Structure
- Composed of two non-covalent lay associated units (alpha and beta) both are required for activity. The Alpha unit contributes to specificity of binding while beta interacts with cytoskeleton
- Binding requires divalent cations (Mg or Mn)
Integrin Function
- No intrinsic enzymatic activity
- Following the binding of integrin to its ligand, the tail of beta subunit binds to proteins talin and alpha actinin
- This then initiates assembly of complex of intracellular attachment proteins that link integrins to actin filaments in cytoskeleton
- Then leads to increases in both intracellular pH and Ca levels and Tyr phosphorylation of proteins
- Binding also initiates assembly of complex of catalytic and regulatory proteins that control gene expression, changes in cytoskeleton and adhesion
- Binding anchors cell to matrix at focal adhesions (bundles of actin filaments -stress fibers- attach to focal adhesions)
Integrins and Disease
- Beta2 subunit forms dimers with different alpha subunits exclusively on surface of WBC’s
- Leukocyte adhesion deficiency - loss of beta2 which eliminates adhesion in WBC’s causing bacterial infections
- Beta3 integrins found in variety of cells, including platelets, and bind several ECM proteins, including fibrinogen
- Glanzmann’s disease - missing or defective beta3 so doesn’t for GPIIb/IIIa which is a receptor for fibrinogen, so platelets can’t bind to fibrinogen to initiate blood clotting, treated with platelet transfusions
- GPIIb/IIIa antagonists used to prevent heart attacks and strokes because they block platelet aggregation and formation of clots
After binding to its ligand, an integrins binds directly to which of the following intracellular proteins?
Talin
Selectins
- cell-surface carbohydrate binding proteins synthesized by endothelial cells, platelets, and WBC
- E or P selectins bind to oligosaccharides of WBC’s, L selectins bind to endothelial cells
- Mediate weak adhesion between WBC and endothelial cells
- Selectins and oligosaccharides are present on both WBC’s and endothelial cells
ICAMs
- intercellular adhesion molecules
- expressed on endothelial cells
- bind to integrins on surface of WBC- heterophilic binding
- mediates stronger interaction which traps WBC’s at sites of inflammation
- specific integrins expressed on WBC’s that recognize ICAM’s
Cell adhesion in inflammation Model
- WBC’s circulate through bloodstream binding weakly to endothelial cells via selectins
- When these cells receive signal from immune system, integrins on surface are activated and bind tightly to endothelial cells lining blood vessel and migrate through endothelial cells to reach site of inflammation
- Integrins of WBC’s often have to be activated, which allows these cells to circulate unimpeded until they are activated by appropriate stimulus
Cadherins
- transmembrane glycoproteins that promote cell adhesion in vertebrate tissues by homophilic binding
- adhesion depends on presence of Ca which prevents proteolysis
- virtually all cells seem to express one or more cadherins
Cadherin structure
- large extracellular region
- highly conserved cytoplasmic domain
- interacts with actin cytoskeleton
Catenins
- alpha and beta catenins
- required for adhesion by cadherins
- alpha catenin links cadherins to actin cytoskeletons
- beta catenin binds to cytoplasmic domain of cadherins and links it to alpha catenin
- p120 binds to cytoplasmic domain of cadherin
Beta catenin and cancer
- Beta catenin is either associated with cadherins or goes into nucleus and binds to transcription factor Tcf-4, which can turn on genes
- Amount of free Beta catenin regulated
- Function depends on Wnt signal
In differentiated cells, no Wnt signal
1) Beta catenin forms complex with APC protein, CK-1 and GSK-3
2) beta catenin is phosphorylase do by CK-1 and GSK-3
3) Beta catenin degraded
4) Tcf protein represses transcription of target genes
In undifferentiated cells, Wnt present
1) binds to frazzled and LRP
2) cytoplasmic protein Dishevelled phosphorylated
3) Leads to phosphorylation of LRP
4) axin associates with receptor, taking CK-1 and GSK-3 with it
5) Beta catenin not degraded
6) Beta catenin translocates to nucleus, forming complex with TcF
7) Complex is transcriptional activator of target genes
APC protein
- product of APC gene
- mutation in APC gene is responsible for FAP which is characterized by hundreds of polyps in colon
- polyps can lead to colon cancer
- mutations in APC can lead to colorectal tumors
- defective APC means beta catenin can’t be degraded.
- target genes include those in cell proliferation (myc, cyclin D1)
- pathway involved in melanomas, liver tumors, and prostate and breast cancer