Uptake kinetics of cell-penetrating peptides.
As our knowledge increases about the diversity in uptake mechanisms displayed by cell-penetrating peptides (CPP), the concept of CPP uptake kinetics becomes increasingly complex. Here, we present three different assays that can be used for studying different kinetic aspects of CPP-mediated delivery: intracellular accumulation and membranolytical effects, intracellular CPP-cargo detachment, and finally a functional readout of a biological action from the delivered cargo. Unlike the traditional end-point measurements that give a static postincubation readout, these assays are all dynamic, real-time, in situ measurements obtained during incubation. A combination of some (or all) of these different assays gives us not only interesting kinetic information about the uptake routes but also provides a simple and valuable methodology for the evaluation of potential drug candidates based on the chemical modification of CPPs by cargo attachment.
Laboratory of Molecular Biotechnology, Institute of Technology, Tartu University, Tartu, Estonia.
Anders
A
Florén
Imre
I
Mäger
Ulo
U
Langel
D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D007700:Kinetics; D021381:Protein Transport; D013050:Spectrometry, Fluorescence; D013696:Temperature
D057846:Cell-Penetrating Peptides; D005456:Fluorescent Dyes