Activated conformations of very late activation integrins detected by a group of antibodies (HUTS) specific for a novel regulatory region (355-425) of the common beta 1 chain.
The very late activation antigens (VLA) or beta 1 integrins mediate cell attachment to different extracellular matrix proteins and intercellular adhesions. The ligand binding activity of these adhesion receptors is not constitutive and can be regulated by temperature, presence of extracellular divalent cations, stimulatory monoclonal antibodies (mAbs), and cellular activation. We have generated three novel mAbs, HUTS-4, HUTS-7, and HUTS-21, recognizing specific epitopes on the common beta 1 subunit (CD29) of VLA integrins whose expression correlates with the ligand binding activity of these heterodimeric glycoproteins. This correlation has been demonstrated for several integrin heterodimers in different cell systems using a variety of extracellular and intracellular stimuli for integrin activation. Thus, the presence of micromolar concentrations of extracellular Mn2+, preincubation with the activating anti-beta 1 mAb TS2/16, and cell treatment with phorbol esters or calcium ionophores, induced the expression of the HUTS beta 1 epitopes on T lymphoblasts. Using a panel of human-mouse beta 1 chimeric molecules, we have mapped these epitopes to the 355-425 sequence of the beta 1 polypeptide. This segment represents therefore a novel regulatory region of beta 1 that is exposed upon integrin activation. Interestingly, binding of HUTS mAbs to partially activated VLA integrins results in maximal activation of these adhesion receptors and enhancement of cell adhesion to beta 1 integrin ligands collagen, laminin, and fibronectin.
Luque
A
A
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense, Madrid, Spain.
Gómez
M
M
Puzon
W
W
Takada
Y
Y
Sánchez-Madrid
F
F
Cabañas
C
C
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003094:Collagen / Q000494:pharmacology; D003110:Colonic Neoplasms; D000939:Epitopes / Q000032:analysis*; D005353:Fibronectins / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D019012:Integrin beta1 / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D007797:Laminin / Q000494:pharmacology; D004915:Leukemia, Erythroblastic, Acute; D008024:Ligands; D008099:Liver / Q000276:immunology; D008168:Lung / Q000276:immunology; D008345:Manganese / Q000494:pharmacology; D018482:Muscle, Skeletal / Q000276:immunology; D011487:Protein Conformation; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005353:Fibronectins; D019012:Integrin beta1; D007797:Laminin; D008024:Ligands; D008345:Manganese; D003094:Collagen