The alpha(4) integrin subunit Tyr(187) has a key role in alpha(4)beta(7)-dependent cell adhesion.
The integrin alpha(4)beta(7) is the cell adhesion receptor for the mucosal vascular addressin MAdCAM-1, and this interaction is dominant in lymphocyte homing to Peyer's patch high endothelial venules, and plays key roles in lymphocyte recruitment at sites of inflammation. To identify alpha(4) subunit amino acids important for alpha(4)beta(7)/MAdCAM-1 interaction, we expressed mutant alpha(4) and wild type beta(7) chains in K562 cells and analyzed the effect of the mutations on cell adhesion to a soluble MAdCAM-1 (sMAdCAM-1-Ig). Transfectants expressing mutated alpha(4) at Tyr(187) displayed a substantial decrease in adhesion to this ligand, which was associated with a reduced alpha(4)beta(7)/sMAdCAM-1-Ig interaction, as determined by soluble binding assays. Addition of Mn(2+) to the adhesion assays did not restore the impaired adhesion. Mutations at alpha(4) Gln(152)Asp(153) also affected transfectant adhesion to sMAdCAM-1-Ig, but did not involve an alteration of alpha(4)beta(7)/MAdCAM-1 binding, and adhesion was restored by Mn(2+). Instead, mutations at alpha(4) Asn(123)Glu(124) did not affect this adhesion. Mutation of alpha(4) Tyr(187) abolished alpha(4)beta(7)-mediated cell adhesion to CS-1/fibronectin, an additional ligand for alpha(4)beta(7), while alpha(4) Gln(152)Asp(153) transfectant mutants showed a reduced adhesion. These results identify alpha(4) Tyr(187) as a key residue during receptor alpha(4)beta(7)/ligand interactions, indicating that it plays important roles in alpha(4)beta(7)-mediated leukocyte adhesion, and provide a potential target for therapeutic intervention in several inflammatory pathologies.
Ruiz-Velasco
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N
Department of Immunology, Centro de Investigaciones Biológicas, Velázquez 144, 28006 Madrid, Spain.
Guerrero-Esteo
M
M
Briskin
M J
MJ
Teixidó
J
J
D015703:Antigens, CD / Q000502:physiology*; D002448:Cell Adhesion; D005353:Fibronectins / Q000502:physiology; D006801:Humans; D039441:Integrin alpha4; D016023:Integrins / Q000502:physiology*; D020014:K562 Cells; D016297:Mutagenesis, Site-Directed; D014162:Transfection; D014443:Tyrosine; D019010:Vascular Cell Adhesion Molecule-1 / Q000502:physiology
D015703:Antigens, CD; D005353:Fibronectins; D016023:Integrins; D019010:Vascular Cell Adhesion Molecule-1; C071777:integrin alpha4beta7; D039441:Integrin alpha4; D014443:Tyrosine