Back to Search
Start Over
A direct binding assay for the vascular cell adhesion molecule-1 (VCAM1) interaction with alpha 4 integrins.
- Source :
-
Cell adhesion and communication [Cell Adhes Commun] 1995 Dec; Vol. 3 (5), pp. 385-97. - Publication Year :
- 1995
-
Abstract
- Vascular cell adhesion molecule-1 (VCAM1) is a member of the immunoglobulin (Ig) superfamily which interacts with the alpha 4 integrins alpha 4 beta 1 (very late antigen 4: VLA4) and alpha 4 beta 7, which are constitutively expressed on many leukocyte subsets and play a key role in cell trafficking and activation. Using a recombinant VCAM-IgG fusion protein (VCAM-Ig) as a soluble ligand for alpha 4 beta 1 we directly demonstrated by fluorescence analysis that the alpha 4 beta 1 receptor can exist in different affinity states on the cell surface, and that a high affinity state is induced by manganese ions or certain activating anti-beta 1 monoclonal antibodies (Jakubowski et al., 1995b). Here we have extended these observations by developing a rapid and reproducible assay using alkaline phosphatase (AP)-coupled VCAM-Ig (VCAM-Ig-AP) which measures the interaction between VCAM1 and alpha 4 integrins in a microtiter plate format. This assay has allowed us to evaluate directly the effects of metal ions, anti-beta 1 mAbs, and different cell types and species on the VCAM1/alpha 4 integrin interaction. Most importantly, the assay system provides a means to rapidly evaluate alpha 4 integrin-directed inhibitors without the complication of post-ligand binding events inherent in adhesion assays.
- Subjects :
- Animals
Antibodies, Monoclonal metabolism
Binding, Competitive drug effects
Binding, Competitive physiology
Cell Adhesion physiology
Fluorescent Antibody Technique
Humans
Integrin alpha4
Leukemia
Magnesium pharmacology
Mice
Protein Binding physiology
Rats
Tumor Cells, Cultured chemistry
Tumor Cells, Cultured cytology
Tumor Cells, Cultured metabolism
Vascular Cell Adhesion Molecule-1 immunology
Antigens, CD metabolism
Integrins metabolism
Vascular Cell Adhesion Molecule-1 metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1061-5385
- Volume :
- 3
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Cell adhesion and communication
- Publication Type :
- Academic Journal
- Accession number :
- 8640376
- Full Text :
- https://doi.org/10.3109/15419069509081293