Intracellular translocation and differential accumulation of cell-penetrating peptides in bovine spermatozoa: evaluation of efficient delivery vectors that do not compromise human sperm motility

Townley

et al. 2015

Nanomedicine (Lond.)

Section: Sperm Motility and Morphology After Exposure To C105y-functionsupporting

confidence: 89%

mentioning

confidence: 99%

Functionalization of Mesoporous Silica Nanoparticles with a Cell-Penetrating Peptide to Target Mammalian Sperm in vitro

Barkalina

Townley

et al. 2015

Nanomedicine (Lond.)

“…As previously reported (see references in Table 1), all peptides were purified to apparent homogeneity by semi-preparative scale high performance liquid chromatography, and their predicted masses (average M + H + ) were confirmed to an accuracy of ± 1.0 by matrix-assisted laser desorption ionization (MALDI) time of flight mass spectrometry operated in positive ion mode using α-cyano-4-hydroxycinnamic acid (Sigma) as a matrix [6,23,24].…”

Section: Peptide Synthesismentioning

confidence: 80%

“…Polycationic LRRK2-derived sequences were synthesized on a 0.1 mmol scale using a using a Discover SPS Microwave Peptide Synthesizer (CEM Microwave Technology Ltd, Buckingham, UK; [24]). The solid phase was Rink amide 4-methylbenzhydrylamine resin preloaded with the first amino acid (AnaSpec, Inc. Cambridge Bioscience Ltd., Cambridge, UK) and employed an N-α-Fmoc protection strategy with (2-(6-chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU) activation.…”

Section: Peptide Synthesismentioning

confidence: 99%

“…RBL-2H3 cells were cultured in 24-well plates as above and pretreated for 1 h with either TAMRA-Tat (3 μM) or a conjugate of BtnTat plus Texas Red-conjugated Avidin (Avidin-TXR; Life Technologies Ltd., Paisley, UK) in HAMS F12 medium without phenol red (Stratech Scientific Ltd., Newmarket, UK). To ensure Btn-Tat-Avidin-TXR complex formation, 1 μM Btn-Tat was mixed with Avidin-TXR at a 3:1 molar ratio for 1 h prior to incubation with cells [24]. Subsequently, exocytosis was either stimulated with increasing concentrations of MP (0.1-30 μM, for 15 min) or by antigen-antibody-mediated crosslinking of FcɛR1.…”

Section: Regulated Exocytosis Of Translocated Peptide Conjugatesmentioning

confidence: 99%

See 1 more Smart Citation

Cell penetrating peptide-mediated transport enables the regulated secretion of accumulated cargoes from mast cells

Howl

Journal of Controlled Release

2015

The in vivo utility of technologies employing cell penetrating peptides and bioportides may be compromised by the general capacity of polycationic peptides to activate mast cell secretion. Moreover, the same technologies could be exploited in a clinical setting either to directly modulate intrinsic exocytotic mechanisms or to load mast cells with bioactive cargoes. Comparative investigations identified two cell penetrating vectors, Tat and C105Y, which readily translocate into mast cells without inducing receptor-independent exocytosis. Efficient Tat transduction also enabled the intracellular delivery and accumulation of cargoes within discrete intracellular compartments. A tetramethylrhodamine-Tat conjugate is effectively translocated into the secretory lysosomes of RBL-2H3 cells. In contract, the intracellular delivery of avidin, as a non-covalent complex with a biotinylated Tat vector, is also efficient but the protein is predominantly accumulated outside of secretory lysosomes. Significantly, both cargoes can be subsequently released following mast cell stimulation either by mastoparan, a wasp venom secretagogue, or by the physiological mechanism of antigen-induced aggregation of high affinity IgE receptors. These studies indicate that mast cells could be exploited to direct the delivery of bioactive agents to disease sites as an innovative cell-mediated therapy.

Bioportides: Bioactive cell‐penetrating peptides that modulate cellular dynamics

Lukanowska

Howl

2013

Biotechnology Journal

The study and exploitation of cell-penetrating peptides (CPPs) now extends into a third exciting decade. Pharmacokinetic modulators, including the more common sequences Tat, penetratin and transportan-10, markedly enhance the intracellular delivery of small drugs, peptides, oligonucleotides and proteins. We introduced the term bioportide to distinguish cell penetrant peptides with intrinsic bioactivities from more typically inert CPP vectors. Our first examples included rhegnylogically organised bioportides, monomeric peptides presenting pharmacophores for both cellular internalization and bioactivity discontinuously distributed within the primary sequence. However, it is conceptually expedient to employ the same terminology to encompass sychnologic bioportides that comprise an inert CPP vector conjugated to an otherwise impermeable bioactive peptide. In such cases the CPP provides an obvious address function whilst the bioactive cargo, often a protein mimetic sequence, is the message. Additional targeting sequences, usually added as chimeric extensions, can also be accommodated within the design of CPPs and bioportides to enable cell- and tissue-selective targeting. Thus, the identification and exploitation of bioportides provides further scope to employ CPPs as research tools, diagnostics and therapeutics spanning a range of pathologies.