Abstract
Peptides, especially intracellular functional peptides that can play a particular role inside a cell, have attracted attention as promising materials to control cell fate. However, hydrophilic materials like peptides are difficult for cells to internalize. Therefore, the screening and design of intracellular functional peptides are more difficult than that of extracellular ones. An effective high-throughput screening system for intracellular functional peptides has not been reported. Here, we demonstrate a novel peptide array system for screening intracellular functional peptides, in which both cell-penetrating peptide (CPP) domain and photo-cleavable linkers are used. By using this screening system, we determined how the cellular uptake properties of CPP-conjugated peptides varied depending on the properties of the conjugated peptides. We found that the internalization ability of CPP-conjugated peptides varied greatly depending on the property of the conjugated peptides, and anionic peptides drastically decreased the uptake ability. We summarized our data in a scatter diagram that plots hydrophobicity versus isoelectric point (pI) of conjugated peptides. These results define a peptide library suitable for screening of intracellular functional peptides. Thus, our system, including the diagram, is a promising tool for searching biological active molecules such as peptide-based drugs.
Highlights
Showing both these activities, an effective screening system for intracellular functional peptides has been aggressively pursued in recent years
As it is difficult to identify intracellular functional peptides by using existing technologies, we investigated the possibility of constructing a novel screening system for intracellular functional peptides
We describe a novel screening system for intracellular functional peptides, in which the previously reported photo-cleavable peptide array[27] is combined with Cell-penetrating peptides (CPPs) (Fig. 1)
Summary
Showing both these activities, an effective screening system for intracellular functional peptides has been aggressively pursued in recent years. We describe a novel screening system for intracellular functional peptides, in which the previously reported photo-cleavable peptide array[27] is combined with CPPs (Fig. 1). The functions of CPPs (e.g., cellular uptake, cytotoxicity) were investigated using a photo-cleavable linker[29].
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