Cell-penetrating peptides (CPPs) are small peptides with a highcontent of basic amino acid residues. They possess the ability to translocate through the plasma membrane and facilitate exogenous cargodelivery into living cells. In this chapter, we demonstrate that argininerichCPPs are able to not only traverse cellular membranes by themselves,but also carry macromolecules into human A549 lung carcinoma cells inmixed covalent and noncovalent manners. This special macromoleculardelivery system was named as mixed covalent and noncovalent proteintransductions (CNPT). We found that cells treated with nona-arginine(R9)-red fluorescent protein (RFP) fusion protein mixed with greenfluorescent protein (GFP), referred to as R9-RFP/GFP complexes, exhibitboth red and green fluorescent images. Cells treated with R9-GFP fusionprotein mixed with RFP, denoted as R9-GFP/RFP complexes, emittedgreen and red fluorescence, vice versa. Furthermore, mechanistic studiesrevealed that the cellular uptake mechanism of CNPT may involve acombination of multiple internalization pathways. Therefore, applicationsof this binary CNPT system may provide an efficient tool for delivery ofmultiple proteins in bioscience and clinical research.
Date:
2013-05
Relation:
Macromolecular Chemistry: New Research. 2013 May 17:69-82.
