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Related Concept Videos

Integrins01:10

Integrins

Animal and protozoan cells do not have cell walls to help maintain shape and provide structural stability. Instead, these eukaryotic cells secrete a sticky mass of carbohydrates and proteins into the spaces between adjacent cells. This network of proteins and molecules is called an extracellular matrix or ECM.
Some ECM proteins assemble into a basement membrane to which the remaining components adhere. Proteoglycans typically form the bulk of the ECM while fibrous proteins, like collagen,...
Intracellular Signaling Affects Focal Adhesions01:17

Intracellular Signaling Affects Focal Adhesions

Integrins act both as extracellular input receivers and as intracellular processing activators. As their name suggests, integrins are entirely integrated into the membrane structure. Their hydrophobic membrane-spanning regions interact with the phospholipid bilayer's hydrophobic region. These membrane receptors provide extracellular attachment sites for effectors like hormones and growth factors. They activate intracellular response cascades when their effectors are bound and active.
Some...
Activation of Integrins01:15

Activation of Integrins

Integrins bind ligands and transmit information from outside the cell to inside or vice-versa through an "outside-in signaling" or "inside-out signaling."
In "outside-in signaling," external factors in the extracellular space bind to exposed ligand binding sites on integrins. This causes the inactive protein to undergo a conformational change to become active. Integrins are often clustered on the cell membrane. Repetitive and regularly spaced ligand binding events provide an effective stimulus.
Immunoglobulin-like Cell Adhesion Molecules01:31

Immunoglobulin-like Cell Adhesion Molecules

Immunoglobulin-like cell adhesion molecules or Ig-CAMs are a versatile group of cell surface glycoproteins belonging to the immunoglobulin protein superfamily. Ig-CAMs possess the characteristic immunoglobulin protein domains and other domains such as the fibronectin type III domain. The Ig domains are glycosylated to varying degrees in different Ig-CAMs.
Ig-CAMs exhibit either homophilic binding (to other Ig-CAMs) or heterophilic binding (to other ligands such as integrins). While most Ig-CAMs...
Selectins01:25

Selectins

Cell adhesion is  an essential aspect of multicellularity. While stable cell interactions usually occur between cells of the same type, transient cell interactions occur between cells of different tissue types, such as between neutrophils and endothelial cells. Selectins are one class of cell adhesion molecules (CAMs) that bind carbohydrate ligands to form transient cell adhesion. They are rod-like proteins with a long extracellular part of variable length ending with the lectin domain, which...
Laminins are the Adhesive Proteins of Basal Lamina00:55

Laminins are the Adhesive Proteins of Basal Lamina

Laminins are heterotrimeric proteins with high molecular mass found in the extracellular matrix. Each laminin molecule is composed of three chains, viz. alpha, beta, and gamma, coded by five, four, and three paralogous genes, respectively. Laminins are categories based on the compositions of the three chains.
In humans, the five forms of alpha chains are LAMA 1, LAMA 2, LAMA 3, LAMA 4, and LAMA 5. The four forms of beta chains are LAMB 1, LAMB 2, LAMB 3, and LAMB 4. The three forms of gamma...

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Ligand Nano-cluster Arrays in a Supported Lipid Bilayer
10:34

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Published on: April 23, 2017

Self-assembling multimeric integrin alpha5beta1 ligands for cell attachment and spreading.

M Kreiner1, Z Li, J Beattie

  • 1Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 27 Taylor Street, Glasgow, UK.

Protein Engineering, Design & Selection : PEDS
|June 3, 2008
PubMed
Summary
This summary is machine-generated.

Engineered protein multimers enhance cell adhesion by presenting clustered arginine-glycine-aspartic acid (RGD) ligands and synergy sites for integrin alpha5beta1 binding. Higher-order structures (tetramers) promoted significantly greater fibroblast spreading on modified surfaces.

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Protein Engineering

Background:

  • Cell adhesion is crucial for tissue engineering and regenerative medicine.
  • Integrin alpha5beta1 binding requires both arginine-glycine-aspartic acid (RGD) motifs and a synergy site on fibronectin type III domains.
  • Current methods for enhancing cell adhesion often lack precise control over ligand presentation.

Purpose of the Study:

  • To design and create novel multimeric protein ligands that specifically bind integrin alpha5beta1.
  • To investigate the effect of ligand valency on cell adhesion and spreading.
  • To develop a method for orienting these ligands on surfaces for improved cell interactions.

Main Methods:

  • Construction and expression of soluble protein chimeras containing fibronectin type III domains (9th-10th) and self-assembling coiled-coil domains.
  • Mutation of leucine zipper domains to create dimeric, trimeric, and tetrameric coiled coils.
  • Surface immobilization of chimeras via C-terminal cysteine anchoring to streptavidin-coated surfaces.
  • Characterization of multimerization and secondary structure using size-exclusion chromatography and circular dichroism.
  • Assessment of integrin alpha5beta1 binding using surface plasmon resonance.
  • Evaluation of fibroblast spreading on derivatized surfaces.

Main Results:

  • Successfully designed and expressed soluble protein chimeras that self-assemble into defined dimers, trimers, and tetramers.
  • Demonstrated specific binding of these chimeras to integrin alpha5beta1.
  • Quantified increased fibroblast spreading on surfaces functionalized with higher-order multimer ligands (tetramer > trimer > dimer).
  • Confirmed defined secondary structures and successful surface anchoring of the protein chimeras.

Conclusions:

  • Novel polyvalent integrin alpha5beta1 ligands were developed through protein engineering.
  • The valency of these ligands significantly influences fibroblast adhesion and spreading.
  • These engineered ligands offer a versatile platform for modifying biomaterial surfaces to enhance cell adhesion in vitro.