Protein delivery by the cell-penetrating peptide YTA2.
In most cases, the transport of cell-penetrating peptide (CPP) with a cargo molecule over the plasma membrane requires a cross-linking of the cargo molecule to the peptide. Lately, a method of cargo delivery, coincubation with CPP, has been applied. We have studied uptake and toxicity of the CPP, YTA2, in the Bowes human melanoma cell line and human MDA-MB-231 breast cancer cell line and compared the results with known cell-penetrating peptides. The results show that fluoresceinyl YTA2 is taken up by the Bowes cells with 3.23 nmol/mg protein and shows low membrane toxicity to the cells with an EC50 of 60 microM. Furthermore, we show that YTA2 is capable of delivering cargo proteins, such as beta-galactosidase and tetramethyl rhodamine iso-thiocyanate (TRITC) labeled streptavidin into cells by coincubation. The delivery of TRITC-labeled streptavidin was quantified to 42.4 pmol streptavidin/mg protein. The delivery of proteins into the cells by mere coincubation is an advantage, since the chemical coupling between the CPP and the cargo molecule, which adds time-consuming synthesis and purification steps, can be omitted. In addition, the flexibility in CPP cargo delivery is increased.
Myrberg
Helena
H
Department of Neurochemistry, Stockholm University, Svante Arrhenius v. 21A, SE-106 91 Stockholm, Sweden. helenam@neurochem.su.se
Lindgren
Maria
M
Langel
Ulo
U
D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005456:Fluorescent Dyes; D006801:Humans; D006836:Hydro-Lyases / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology* / Q000633:toxicity
D005456:Fluorescent Dyes; D010455:Peptides; C508980:YTA2 peptide; D000255:Adenosine Triphosphate; D006836:Hydro-Lyases; C045581:lactate dehydratase