title: Discovery of a non-cationic cell penetrating peptide derived from membrane-interacting human proteins and its potential as a protein delivery carrier.
abstract: Cell penetrating peptides (CPPs) are peptides that can be translocated into cells and used as a carrier platform for the intracellular uptake of cargo molecules. Subject to the source of CPP sequences and their positively charged nature, the cytotoxicity and immunogenicity of conventional CPPs needs to be optimized to expand their utility for biomedical applications. In addition to these safety issues, the stability of CPPs needs to be addressed since their positively charged residues are prone to interact with the biological milieu. As an effort to overcome these limitations of the current CPP technology, we isolated CPP candidate sequences and synthesized peptides from twelve isoforms of annexin, a family of membrane-interacting human proteins. The candidate screen returned a CPP rich in hydrophobic residues that showed more efficient cellular uptake than TAT-CPP. We then investigated the uptake mechanism, subcellular localization, and biophysical properties of the newly found CPP, verifying low cytotoxicity, long-term serum stability, and non-immunogenicity. Finally, model proteins conjugated to this peptide were successfully delivered into mammalian cells both in vitro and in vivo, indicating a potential use of the peptide as a carrier for the delivery of macromolecular cargos.
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology (KIST), Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Hyo
authors.initials: H
authors.lastname: Young Kim
authors.affiliation: Laboratory of Theriogenology, Department of Veterinary Clinical Science, College of Veterinary Medicine and BK21 PLUS Program for Creative Veterinary Science Research, Seoul National University.
authors.forename: Soo
authors.initials: S
authors.lastname: Young Yum
authors.affiliation: 1] Laboratory of Theriogenology, Department of Veterinary Clinical Science, College of Veterinary Medicine and BK21 PLUS Program for Creative Veterinary Science Research, Seoul National University [2] Emergence Center for Food-Medicine Personalized Therapy System, Advanced Institutes of Convergence Technology, Seoul National University, Gyeonggi-do 443-270, Korea.
authors.forename: Goo
authors.initials: G
authors.lastname: Jang
authors.affiliation: 1] Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology (KIST), Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea [2] Department of Biological Chemistry, Korea University of Science and Technology (UST), KIST campus, Republic of Korea.
authors.forename: Dae-Ro
authors.initials: DR
authors.lastname: Ahn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins / Q000737:chemistry; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000494:pharmacology*; D002942:Circular Dichroism; D004337:Drug Carriers; D016503:Drug Delivery Systems; D015195:Drug Design; D004705:Endocytosis / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D020033:Protein Isoforms / Q000378:metabolism; D055550:Protein Stability; D021381:Protein Transport; D000067996:RAW 264.7 Cells; D013347:Subcellular Fractions / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D017302:Annexins; D002412:Cations; D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; D008565:Membrane Proteins; D020033:Protein Isoforms; D001616:beta-Galactosidase