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  Integrin reconstituted in GUVs: a biomimetic system to study initial steps of cell spreading

Streicher, P., Nassoy, P., Bärmann, M., Dif, A., Marchi-Artzner, V., Brochard-Wyart, F., et al. (2009). Integrin reconstituted in GUVs: a biomimetic system to study initial steps of cell spreading. Biochimica et Biophysica Acta (BBA), 1788(10), 2291-2300. doi:10.1016/j.bbamem.2009.07.025.

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BBA_1788_2009_2291.pdf (Any fulltext), 686KB
 
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 Creators:
Streicher, Pia1, Author           
Nassoy, Pierre, Author
Bärmann, Michael, Author
Dif, Aurélien, Author
Marchi-Artzner, Valérie, Author
Brochard-Wyart, Françoise, Author
Spatz, Joachim P.1, 2, Author           
Bassereau, Patricia, Author
Affiliations:
1Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_2364731              
2Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany, ou_persistent22              

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Free keywords: Adhesion; Integrin; Membrane protein reconstitution; Spreading kinetics; RICM; GUV; Fibrinogen; Quantum dots
 Abstract: A novel in vitro membrane system mimicking the first steps of integrin-mediated cell spreading has been developed and characterized. We have reconstituted the transmembrane alpha(IIb)beta(3) integrin into giant unilamellar vesicles (GUVs). The reconstitution process has been validated by analyzing protein incorporation and biological activity by checking the specific interaction of GUVs containing integrin with quantum dots (QD) or surfaces coated with the integrin receptor tri-peptide RGD.(1) The spreading dynamics of integrin-functionalized GUVs onto fibrinogen-coated surfaces has been monitored by Reflection Interference Contrast Microscopy (RICM). Our results are quantitatively consistent with a theoretical model based on a dewetting process coupled to binder diffusion and provide a comprehensive description of the following sequence: i) nucleation and growth of adhesive patches coupled to the diffusion of the adhesive proteins to these adhesive zones ii) fusion of patches and formation of an adhesive ring iii) complete spreading of the GUV by dewetting of the central liquid film from the border to form an adhesive circular patch that is not significantly enriched in integrins, as compared to the unbound membrane. This finding is consistent with the recognized role of the actin cytoskeleton in stabilizing focal complexes and focal adhesions in a cell-extracellular matrix contact. These very large unilamellar integrin-containing vesicles provide a unique artificial system, which could be further developed towards realistic cell mimic and used to study the complexity of integrin-mediated cell spreading.

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Language(s): eng - English
 Dates: 2009-07-102009-01-202009-07-272009-08-072009-10-01
 Publication Status: Issued
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

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Title: Biochimica et Biophysica Acta (BBA)
  Other : Biochimica et Biophysica Acta (BBA) - Biomembranes
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 1788 (10) Sequence Number: - Start / End Page: 2291 - 2300 Identifier: Other: 1879-2642
CoNE: https://pure.mpg.de/cone/journals/resource/18792642