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Use of GPI‐anchored proteins to study biomolecular interactions by surface plasmon resonance
Author(s) -
Harrison Patrick T,
Campbell Ian W,
Allen Janet M
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00027-1
Subject(s) - surface plasmon resonance , receptor , biophysics , ligand (biochemistry) , biosensor , cell surface receptor , chemistry , receptor–ligand kinetics , cell membrane , membrane protein , cell , membrane , biochemistry , nanotechnology , biology , materials science , nanoparticle
Surface plasmon resonance is a powerful tool to examine the kinetics of cell surface receptor‐ligand interactions and requires only small amounts of protein. For these studies, one component is required in highly purified form to be coupled to the biosensor surface. The second component does not need to be purified. The human high affinity receptor for immunoglobulin G, FcγRI, presents a problem as the receptor itself cannot readily be produced in large amounts for purification and, as there are eight potential ligands for the receptor (human IgG1–4 and mouse IgG1, 2a, 2b and 3), it is difficult to immobilise the ligand. Using a previously established method for generating GPI‐anchored proteins, we have produced and captured a soluble version of FcγRI and shown that it retains its affinity for human IgG1 and specificity for the different IgG subclasses. In addition, we also produced and captured a GPI‐anchored version of the cell adhesion molecule CD2. This system circumvents the need for extensive receptor purification and is very rapid as solubilised receptors can be transferred from the cell surface to the sensor chip in 2 h. This system may be generally applicable for biosensor studies to other type I membrane proteins, and/or naturally occurring GPI‐anchored proteins, especially where the interaction between a ligand and a panel of variant receptors is to be studied.