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A reversible cell penetrating peptide-cargo linkage allows dissection of cell penetrating peptide- and cargo-dependent effects on internalization and identifies new functionalities of putative endolytic peptides

Front Pharmacol .. 2022-11; 
Daniel P Morris, Lucy C Snipes, Stephanie A Hill, Michael M Woods, Maria M Mbugua, Lydia R Wade, Jonathan L McMurry
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Gene Synthesis For example, pJM161, encoding TAT-naked mole rat calmodulin (TATNMR-CaM) consisted of an E. coli-optimized synthetic gene (GenScript, Piscataway, NJ, United States) cloned into NdeI and BamHI sites in pET19b with an in-frame stop codon prior to the BamHI site. Get A Quote

摘要

Cell penetrating peptides (CPPs) are a promising technology for therapeutic delivery of macromolecular cargos. CPPs have generally used covalent linkages to cargo, ensuring a common fate as one molecule. Conversely, our CPP-adaptor, TAT-CaM, noncovalently binds calmodulin binding sequence (CBS)-containing cargos in calcium rich media then dissociates in the calcium-poor endosomal environment following internalization, enhancing endosomal escape relative to standard CPPs. In this study, we report cell entry of positively charged protein cargos that were not increased by TAT-CaM while cargos based on the negatively charged maltose binding protein (MBP) displayed little intrinsic internalization but were internali... More

关键词

calmodulin (CAM); cell-penetrating peptides; endocytosis; endolytic peptides; protein transduction domains (PTD).