Proceedings book download - 5Z.com

Table 1. Apoptogenic and translocation profilesPeptideSequenceApoptogenicPotency(LD 50) aTranslocationEfficacy bIntracellularTarget (colocalizationcoefficient) cCyt c 77-101H-GTKMIFVGIKKKEERADLIAYLKKA80.63 M105.9 + 0.42EndoplasmicReticulum(0.996 + 0.02)NH 2CH 3 CO-NFKFGLSS(Ahx)GTKMIFVGIKKKEERADLIAYLKKAAc-Nup153-Cyt c0.73 M408.5 + 15.8Nucleoporin153(0.931 + 0.02)NH 2CH 3 CO-GFSNSFKLN(Ahx)GTKMIFVGIKKKEERADLIAYLKKAAc-ScrNup153-Cyt cNot toxic412.5 + 5.20CytoplasmicNH 2a LD 50 values indicate the peptide concentration required to reduce U373MG cell viabilityby 50%. Values were obtained using the MTT conversion assay [5] and % viabilityexpressed as a % of those cells treated with medium alone. Cells were exposed to peptidesfor 24 hours and LD 50 values were calculated from 3 independent experiments performedin sextruplicate.b Translocation efficacies were measured using rhodamine-labelled peptides. U373MGcells were incubated with peptides (5 M) for 1 hour. Thereafter, cell lysates were analyzedusing fluorescence spectroscopy and results expressed as fold uptake compared tountreated cells. Data are from 3 independent experiments performed in triplicate.c Intracellular co-localization analysis was carried out using live confocal cell imaging ofU3773MG cells treated with rhodamine-labelled peptides (3 M) for 1 hour. Colocalizationcoefficients were calculated using the Carl Zeiss quantitative co-localizationanalysis software. Coefficient values (overlap coefficient after Manders) are between 0and 1, where a value of 1 indicates that all pixels are co-localized, whilst a value of 0denotes no co-localization.AcknowledgmentsWe would like to thank Imre Mäger at the Laboratory of Molecular Biotechnology, Institute ofTechnology, Tartu University, Estonia and Professor Ülo Langel at the Department of Neurochemistry,Stockholm University, Sweden for their assistance in the synthesis of Fluo-Nup153-M918.References1. Howl, J., Jones, S. Int. J. Pept. Res. Ther. 14, 359-366 (2008).2. Johansson, H.J., et al. Mol. Ther. 16, 115-123 (2008).3. Östlund, P., et al. Int. J. Pept. Res. Ther. 11, 237-247 (2005).4. Hällbrink, M. Int. J. Pept. Res. Ther. 11, 249-259 (2005).5. Carmichael, J., et al. Cancer Research 47, 936-942 (1987).417