Title :
Kinetics of LFA-1 binding to ICAM-1 studied in a cell-free system
Author :
Sarantos, M.R. ; Lum, A.F.H. ; Staunton, D.E. ; Simon, S.I.
Author_Institution :
Dept. of Biomedical Eng., California Univ., Davis, CA, USA
Abstract :
Neutrophil capture on inflamed endothelium is controlled by dynamic regulation of the integrin CD11a/CD18 (LFA-1). Small molecules, antibodies, and certain divalent cations binding to specific epitopes on the integrin are able to stabilize either a closed (low affinity) or open (high affinity) state. To determine the relationship between LFA-1 conformation and affinity for ICAM-1 we assembled a cell-free system consisting of CD11a/CD18 heterodimer adhered to latex microspheres. The kinetics of dimeric ICAM-1 binding to the LFA-1 on the microspheres was measured via flow cytometry and a real time conformational shift into a lower affinity state was observed by addition of a small molecule inhibitor.
Keywords :
adhesion; blood; cellular biophysics; molecular biophysics; molecular configurations; ICAM-1; LFA-1; LFA-1 conformation; adhesion; antibodies; binding affinity; binding kinetics; cell-free system; divalent cations; flow cytometry; inflamed endothelium; integrin CD11a/CD18 (LFA-I); neutrophil capture; real time conformational shift; small molecule inhibitor; Adhesives; Assembly systems; Biomembranes; Blood flow; Blood vessels; Fluid flow measurement; Fluorescence; Inhibitors; Kinetic theory; Wounds; Inflammation; integrin conformation; neutrophil adhesion;
Conference_Titel :
Engineering in Medicine and Biology Society, 2004. IEMBS '04. 26th Annual International Conference of the IEEE
Conference_Location :
San Francisco, CA
Print_ISBN :
0-7803-8439-3
DOI :
10.1109/IEMBS.2004.1404375
