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Fig. 2. Staining of SHSY-5Y cells incubated with R2 using TRITC-avidin (plate c), PSD-95using FITC-anti-rabbit and PSD-95 rabbit antibody (Abnova) (plate b), showedco-localisation (plate a).The peptide R2 was readily internalized, showing that the hexa-D-Arg sequence acted as aneffective cell penetration peptide. The biotin moiety in R2 co-stained with an antibodybinding to PSD-95 (Figure 2), indicating that the PDZ domain in PSD-95 was targeted byR2. SHSY-5Y cells challenged with 2mM glutamate were effectively protected against celldeath by 2μM, 20μM and 200μM of the N-acetylated and non-acetylated forms of R2 inhuman non-differentiated SHSY-5Y neuroblastoma cells (Figure 3). Further modificationsare required, however, to increase affinity and give rise to an effective therapeutic that willcross the blood-brain barrier.% cell death.706050403020100-10-20-300 220 200Concentration of PDZ-binding peptides (micromolar)Acetyl R2NonAcR2H2Fig. 3. Protection against cell death of glutamic acid (2mM) treated SHSY-5Y cells.AcknowledgmentsWe acknowledge the awarding of a Biochemical Society (UK) travel grant to Kate Duberley.References1. Garside, M.L., Turner, P.R., Austen, B., Strehler, E.E., Beesley, P.W., Empson, R.M. Neuroscience162, 383-395 (2009).2. Piserchio, A., Salinas, G.D., Li, T., Marshall, J., Spaller, M.R., Mierke, D.F. Chem.Biol. 11, 469-473 (2004).545