Mapping HA-tagged protein at the surface of living cells by atomic force microscopy

摘要: Single-molecule force spectroscopy using atomic microscopy (AFM) is more and used to detect map receptors, enzymes, adhesins, or any other molecules at the surface of living cells. To be specific, this technique requires antibodies ligands covalently attached AFM tip that can specifically interact with protein interest. Unfortunately, specific are usually lacking (low affinity specificity) expensive produce (monoclonal antibodies). An alternative strategy tag interest a peptide recognized high specificity commercially available antibodies. In context, we chose work human influenza hemagglutinin (HA) (YPYDVPDYA) labeled two proteins: linked cell wall 12 (Ccw12) involved in remodeling yeast Saccharomyces cerevisiae β2-adrenergic receptor (β2-AR), G protein-coupled (GPCR) higher eukaryotes. We first described interaction between HA antibodies, immobilized on tips, epitopes, epoxy glass slides. Using our system, then investigated distribution Ccw12 proteins over S. cerevisiae. were able find tagged mating yeasts, projections. Finally, could unfold multimers β2-AR from membrane transfected chinese hamster ovary This result agreement GPCR oligomerization membranes opens door study influence process.