Requirement of open headpiece conformation for activation of leukocyte integrin alphaXbeta2.
Negative stain electron microscopy (EM) and adhesion assays show that alpha(X)beta(2) integrin activation requires headpiece opening as well as extension. An extension-inducing Fab to the beta(2) leg, in combination with representative activating and inhibitory Fabs, were examined for effect on the equilibrium between the open and closed headpiece conformations. The two activating Fabs stabilized the open headpiece conformation. Conversely, two different inhibitory Fabs stabilized the closed headpiece conformation. Adhesion assays revealed that alpha(X)beta(2) in the extended-open headpiece conformation had high affinity for ligand, whereas both the bent conformation and the extended-closed headpiece conformation represented the low affinity state. Intermediate integrin affinity appears to result not from a single conformational state, but from a mixture of equilibrating conformational states.
Chen
Xing
X
Department of Pathology, Harvard Medical School, Immune Disease Institute, Children's Hospital Boston, Boston, MA 02115, USA.
Xie
Can
C
Nishida
Noritaka
N
Li
Zongli
Z
Walz
Thomas
T
Springer
Timothy A
TA
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D001665:Binding Sites; D002448:Cell Adhesion / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000378:metabolism; D016167:Integrin alphaXbeta2 / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D007962:Leukocytes / Q000737:chemistry* / Q000276:immunology; D008024:Ligands; D008854:Microscopy, Electron; D008958:Models, Molecular; D011485:Protein Binding / Q000276:immunology; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D016167:Integrin alphaXbeta2; D008024:Ligands