Designing cell-permeant phosphopeptides to modulate intracellular signaling pathways

Dunican, Dara J.; Doherty, Patrick

doi:10.1002/1097-0282(2001)60:1<45::aid-bip1003>3.0.co;2-9

Cited by 65 publications

(23 citation statements)

References 99 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Development 134 (24) reports of rapid uptake of TAT and other protein-transductiondomain-tagged peptides (Dunican and Doherty, 2001;Green et al, 2003). The distribution of the labelled CTN appeared to be Gaussian along the z-axis for somata, neurites and growth cones (see Fig.…”

Section: Research Articlementioning

confidence: 99%

Neuronal calcium sensor-1 modulation of optimal calcium level for neurite outgrowth

et al. 2007

View full text Add to dashboard Cite

Neurite extension and branching are affected by activity-dependent modulation of intracellular Ca 2+ , such that an optimal window of [Ca 2+ ] i is required for outgrowth. Our understanding of the molecular mechanisms regulating this optimal [Ca 2+ ] i remains unclear. Taking advantage of the large growth cone size of cultured primary neurons from pond snail Lymnaea stagnalis combined with dsRNA knockdown, we show that neuronal calcium sensor-1 (NCS-1) regulates neurite extension and branching, and activitydependent Ca 2+ signals in growth cones. An NCS-1 C-terminal peptide enhances only neurite branching and moderately reduces the Ca 2+ signal in growth cones compared with dsRNA knockdown. Our findings suggest that at least two separate structural domains in NCS-1 independently regulate Ca 2+ influx and neurite outgrowth, with the C-terminus specifically affecting branching. We describe a model in which NCS-1 regulates cytosolic Ca 2+ around the optimal window level to differentially control neurite extension and branching.

show abstract

Section: Research Articlementioning

confidence: 99%

Neuronal calcium sensor-1 modulation of optimal calcium level for neurite outgrowth

et al. 2007

View full text Add to dashboard Cite

show abstract

“…The peptides correspond to amino acids 23-40 of RhoA (the class III binding domain), amino acids 75-92 of RhoA (the class I binding domain), amino acids 17-32 of Rac 1, and amino acids 17-32 of Cdc42 (Rac and Cdc42 CRIB domains). Each peptide was tagged with the TAT internalisation sequence (GRKKRRQRRRPPQC) at its C-terminus to facilitate transport into the cytoplasm (Dunican and Doherty, 2001). The peptides were dissolved directly into culture medium.…”

Section: Pharmacological Reagentsmentioning

confidence: 99%

Temporally and spatially coordinated roles for Rho, Rac, Cdc42 and their effectors in growth cone guidance by a physiological electric field

Rajnicek

Foubister

McCaig

2006

Journal of Cell Science

101

View full text Add to dashboard Cite

Although it is known that neuronal growth cones migrate towards the cathode of an applied direct current (DC) electric field (EF), resembling the EF present in the developing nervous system, the underlying mechanism remains unclear. Here, we demonstrate temporally and spatially coordinated roles for the GTPases Rac, Cdc42 and Rho and their effectors. Growth cones of cultured Xenopus embryonic spinal neurons turned towards the cathode but collective inhibition of Rho, Rac and Cdc42 attenuated turning. Selective inhibition of Rho, Cdc42 or Rac signalling revealed temporally distinct roles in steering by an electrical gradient. Rho, Rac and Cdc42 are each essential for turning within the initial 2 hours (early phase). Later, Rho and Cdc42 signals remain important but Rac signalling dominates. The EF increased Rho immunofluorescence anodally. This correlated spatially with collapsed growth cone morphology and reduced anodal migration rates, which were restored by Rho inhibition. These data suggest that anodally increased Rho activity induces local cytoskeletal collapse, biasing growth cone advance cathodally. Collapse might be mediated by the Rho effectors p160 Rho kinase and myosin light chain kinase since their inhibition attenuated early turning. Inhibitors of phosphoinositide 3-kinase, MEK1/2 or p38 mitogen-activated protein kinase (MAPK) did not affect turning behaviour, eliminating them mechanistically. We propose a mechanism whereby Rac and Cdc42 activities dominate cathodally and Rho activity dominates anodally to steer growth cones towards the cathode. The interaction between Rho GTPases, the cytoskeleton and growth cone dynamics is explored in the companion paper published in this issue. Our results complement studies of growth cone guidance by diffusible chemical gradients and suggest that growth cones might interpret these co-existing guidance cues selectively.

show abstract

“…However, it has been reported that the use of short (10 -30 amino acids) cationic peptides may provide a solution by enhancing the intracellular delivery of such intractable molecules, both in vitro and in vivo (see reviews in Refs. [1][2][3].…”

mentioning

confidence: 99%

Studies on the Internalization Mechanism of Cationic Cell-penetrating Peptides

Drin

Cottin

Blanc

et al. 2003

Journal of Biological Chemistry

498

397

View full text Add to dashboard Cite

A great deal of data has been amassed suggesting that cationic peptides are able to translocate into eucaryotic cells in a temperature-independent manner. Although such peptides are widely used to promote the intracellular delivery of bioactive molecules, the mechanism by which this cell-penetrating activity occurs still remains unclear. Here, we present an in vitro study of the cellular uptake of peptides, originally deriving from protegrin (the SynB peptide vectors), that have also been shown to enhance the transport of drugs across the blood-brain barrier. In parallel, we have examined the internalization process of two lipid-interacting peptides, SynB5 and pAntp-(43-58), the latter corresponding to the translocating segment of the Antennapedia homeodomain. We report a quantitative study of the time-and dose-dependence of internalization and demonstrate that these peptides accumulate inside vesicular structures. Furthermore, we have examined the role of endocytotic pathways in this process using a variety of metabolic and endocytosis inhibitors. We show that the internalization of these peptides is a temperatureand energy-dependent process and that endosomal transport is a key component of the mechanism. Altogether, our results suggest that SynB and pAntp-(43-58) peptides penetrate into cells by an adsorptive-mediated endocytosis process rather than temperature-independent translocation.

show abstract

Designing cell-permeant phosphopeptides to modulate intracellular signaling pathways

Cited by 65 publications

References 99 publications

Neuronal calcium sensor-1 modulation of optimal calcium level for neurite outgrowth

Neuronal calcium sensor-1 modulation of optimal calcium level for neurite outgrowth

Temporally and spatially coordinated roles for Rho, Rac, Cdc42 and their effectors in growth cone guidance by a physiological electric field

Studies on the Internalization Mechanism of Cationic Cell-penetrating Peptides

Contact Info

Product

Resources

About