Kinetic analysis of cellular internalization and expulsion of unstructured D-chirality cell penetrating peptides |
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Authors: | Manibarathi Vaithiyanathan Hannah C. Hymel Nora Safa Olivia M. Sanchez Jacob H. Pettigrew Cole S. Kirkpatrick Ted J. Gauthier Adam T. Melvin |
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Affiliation: | 1. Cain Department of Chemical Engineering, Louisiana State University, Louisiana, USA;2. LSU AgCenter Biotechnology Lab, Louisiana State University, Louisiana, USA |
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Abstract: | Most cell penetrating peptides (CPPs) are unstructured and susceptible to proteolytic degradation. One alternative is to incorporate D-chirality amino acids into unstructured CPPs to allow for enhanced uptake and intracellular stability. This work investigates CPP internalization using a series of time, concentration, temperature, and energy dependent studies, resulting in a three-fold increase in uptake and 50-fold increase in stability of D-chirality peptides over L-chirality counterparts. CPP internalization occurred via a combination of direct penetration and endocytosis, with a percentage of internalized CPP expelling from cells in a time-dependent manner. Mechanistic studies identified that cells exported the intact internalized D-chirality CPPs via an exocytosis independent pathway, analogous to a direct penetration method out of the cells. These findings highlight the potential of a D-chirality CPP as bio-vector in therapeutic and biosensing applications, but also identify a new expulsion method suggesting a relationship between uptake kinetics, intracellular stability, and export kinetics. |
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Keywords: | cell penetrating peptide chirality direct penetration export kinetics intracellular stability |
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