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Characterizing membrane clustering of the β2 integrin CR3 using fluorescence resonance energy transfer (FRET)
Author(s) -
Heflin Katie Elizabeth,
O’Brien Xian M.,
Hyun YoungMin,
Kim Minsoo,
Albina Jorge E.,
Reichner Jonathan S.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1122.14
Subject(s) - förster resonance energy transfer , integrin , chemistry , priming (agriculture) , biophysics , integrin alpha m , microbiology and biotechnology , receptor , fluorescence , biochemistry , biology , physics , germination , botany , quantum mechanics
β‐glucan, a molecule found in yeast cell walls, has been implicated in priming the immune system without causing an increase in inflammatory cytokines. Our previous data indicates that in neutrophils the primary receptor for β‐glucan is CR3 (αMβ2), a β2 integrin crucial for leukocyte recruitment to areas of inflammation. We undertook a fluorescence resonance energy transfer (FRET) approach to investigate the mechanism of CR3 priming. CR3 activation, like that of the related integrin αLβ2, may be regulated through the affinity of a single molecule for its ligand and/or the clustering of many receptors. Our lab had previously generated constructs of αM and β2 with a C‐terminal fusions of mCFP and mYFP respectively for FRET investigations of affinity activation. Now we have created a new construct of αM with a C‐terminal fusion of mYFP. Both αM constructs, along with a wild‐type β2, were transfected into K562 and HL‐60 cells. All constructs were validated for function and are currently being used to investigate the clustering patterns of CR3 under previously characterized priming conditions. Research supported by NIHGM016694.