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Integrin and Cell Adhesion Molecules

Volume 757 of the series Methods in Molecular Biology pp 205-214

Date:

Monitoring Integrin Activation by Fluorescence Resonance Energy Transfer

  • Craig T. LefortAffiliated withDepartment of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester
  • , Young-Min HyunAffiliated withDepartment of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester
  • , Minsoo KimAffiliated withDepartment of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester Email author 

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Abstract

Aberrant integrin activation is associated with several immune pathologies. In leukocyte adhesion deficiency (LAD), the absence or inability of β2 integrins to undergo affinity upregulation contributes to recurrent infectious episodes and impaired wound healing, while excessive integrin activity leads to an exaggerated inflammatory response with associated tissue damage. Therefore, integrin activation is an attractive target for immunotherapies, and monitoring the effect of agents on integrin activation is necessary during preclinical drug development. The activation of integrins involves the structural rearrangement of both the extracellular and cytoplasmic domains. Here, we describe methods for monitoring integrin conformational activation using fluorescence resonance energy transfer (FRET).

Key words

FRET Integrin Cell adhesion LFA-1 Mac-1 VLA-4 Fluorescence microscopy Flow cytometry