A single amino acid in the cytoplasmic domain of the beta(2) integrin lymphocyte function-associated antigen-1 regulates avidity-dependent inside-outsignaling
Da. Bleijs et al., A single amino acid in the cytoplasmic domain of the beta(2) integrin lymphocyte function-associated antigen-1 regulates avidity-dependent inside-outsignaling, J BIOL CHEM, 276(13), 2001, pp. 10338-10346
The leukocyte-specific beta (2) integrin lymphocyte function-associated ant
igen-1 (LFA-1) (alpha (L)/beta (2)) mediates activation-dependent adhesion
to intercellular adhesion molecule (ICAM)-1, In leukocytes, LFA-1 requires
activation by intracellular messengers to bind ICAM-1, We observed malfunct
ioning of LFA-1 activation in leukemic T cells and K562-transfected cells.
This defective inside-out integrin activation is only restricted to beta (2
) integrins, since beta (1) integrins expressed in KS62 readily respond to
activation signals, such as phorbol la-myristate 13-acetate, To unravel the
se differences in inside-out signaling between beta (1) and beta (2) integr
ins, we searched for amino acids in the p, cytoplasmic domain that are crit
ical in the activation of LFA-1, We provide evidence that substitution of a
single amino acid (L732R) in the beta (2) cytoplasmic DLRE motif, creating
the DRRE motif, is sufficient to completely restore PMA responsiveness of
LFA-1 expressed in K562. In addition, an intact TTT motif in the C-terminal
domain is necessary for the acquired PMA responsiveness. We observed that
restoration of the PMA response altered neither LFA-1 affinity nor the phos
phorylation status of LFA-1. In contrast, strong differences were observed
in the capacity of LFA-1 to form clusters, which indicates that inside-out
activation of LFA-1 strongly depends on cytoskeletal induced receptor reorg
anization that was induced by activation of the Ca2+-dependent protease cal
pain.