Physical Sciences and Mathematics Commons^™

Effect Of Loading Method On A Peptide Substrate Reporter In Intact Cells [Post-Print], Rahuljeet Chadha, Grigorii Kalminskii, Allison Tierney, Joshua Knopf, Lorena Lazo De La Vega, Berjana Mcelrath, Michelle Kovarik

Faculty Scholarship

Studies of live cells often require loading of exogenous molecules through the cell membrane; however, effects of loading method on experimental results are poorly understood. Therefore, in this work, we compared three methods for loading a fluorescently labeled peptide into cells of the model organism Dictyostelium discoideum. We optimized loading by pinocytosis, electroporation, and myristoylation to maximize cell viability and characterized loading efficiency, localization, and uniformity. We also determined how the loading method affected measurements of enzyme activity on the peptide substrate reporter using capillary electrophoresis. Loading method had a strong effect on the stability and phosphorylation of the …