Cellular Internalization of Enhanced Green Fluorescent Protein Ligated to a Human Calcitonin-Based Carrier Peptide

Machova, Zuzana; Mühle, Christiane; Krauss, Ulrike; Tréhin, Rachel; Koch, Annette; Merkle, Hans P.; Beck‐Sickinger, Annette G.

doi:10.1002/1439-7633(20020703)3:7<672::aid-cbic672>3.0.co;2-d

Cited by 47 publications

(65 citation statements)

References 33 publications

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“…Hence the 9-32 fragment was investigated with respect to its ability to behave as a carrier peptide [20]. The 9-32 fragment of hCT, called hCT(9-32) has been used for transfer of green flurescent protein (GFP) through cellular membranes via a covalent carrier-cargo linkage [104] and also for the delivery of daunorubicine into several cell lines [105]. More recently, another derivative of hCT has been developed in which an NLS has been branched on a lysine side chain.…”

Section: Calcitonin-derived Peptidesmentioning

confidence: 99%

Cell-penetrating peptides: tools for intracellular delivery of therapeutics

Deshayes

Morris

Divita

et al. 2005

CMLS, Cell. Mol. Life Sci.

443

332

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The main problem of therapeutic efficiency lies in the crossing of cellular membranes. Therefore, significant effort is being made to develop agents which can cross these barriers and deliver therapeutic agents into cellular compartments. In recent years, a large amount of data on the use of peptides as delivery agents has accumulated. Several groups have published the first positive results using peptides for the delivery of therapeutic agents in relevant animal models. These peptides, called cell-penetrating peptides (CPPs), are short peptides (fewer than 30 residues) with a net positive charge and acting in a receptor- and energy-independent manner. Here, we give an extensive review of peptide-mediated delivery systems and discuss their applications, with particular focus on the mechanisms leading to cellular internalization.

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Section: Calcitonin-derived Peptidesmentioning

confidence: 99%

Cell-penetrating peptides: tools for intracellular delivery of therapeutics

Deshayes

Morris

Divita

et al. 2005

CMLS, Cell. Mol. Life Sci.

443

332

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“…Previously the metabolic degradation of full-length hCT was studied. The obtained cleavage pattern differed from that of hCT(9-32) since full-length hCT was mainly degraded between Phe 16 -Asn 17 , Lys 18 -Phe 19 and Phe 19 -His 20 [28,34] (Fig. (2)).…”

Section: Metabolic Stability Of Several Hct Derived Sequencesmentioning

confidence: 91%

“…(1)). This branched peptide, namely hCT(9-32)-br, showed noticeably higher uptake activities as the linear analogue and seems to be a promising tool for delivering nucleic acid cargos into the cytoplasm [18,19]. With the help of the side chain complex formation with negatively charged molecules, for instance plasmids or aptamers, can be promoted and the delivery to the nucleus can be ensured.…”

Section: Metabolic Stability Of Several Hct Derived Sequencesmentioning

confidence: 99%

“…Shorter sequences were not capable to cross the cell membrane, whereas longer fragments in fact were found to internalize. The efficiency of translocation visually decreased in the order of hCT(9-32) ≈ hCT(12-32) > hCT(15-32) > hCT (18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32). Thus, it seems that the core unit plays an important role for the formation of the secondary structure.…”

Section: Key Motifs Important For the Trans-location Processmentioning

confidence: 99%

“…Recently, hCT(9-32) and its derivatives were shown to successfully deliver several cargos into cells like the antiproliferative drug daunorubicin [17], plasmid DNA [18], aptamers (unpublished data) and even a high molecular weight molecule as the enhanced green fluorescent protein [19]. Increasing interest is seen, therefore, in the use of hCT derived peptide fragments as delivery vectors.…”

Section: Introductionmentioning

confidence: 99%

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Calcitonin-Derived Carrier Peptides

Neundorf

Ag²

2005

CPD

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The recent discovery of carrier peptides offers new opportunities to translocate several bioactive molecules into the cytoplasm. Previous studies have shown that human calcitonin (hCT) and selected C-terminal sequences translocate in nasal epithelium. Moreover, the hCT(9-32) fragment was found to internalize efficiently a number of substances like fluorophores, nucleic acids or the enhanced green fluorescent protein (EGFP). In order to understand the uptake mechanism interactions of hCT(9-32) with membrane models of different lipid compositions have been investigated. From these studies it was possible to shed light on the conformational state of the peptide in the presence of membrane-like conditions. Further insight into the translocation mechanism was provided by fluorescence microscopy of truncated sequences of hCT that were shown to penetrate the plasma membrane and to distribute in a sectoral, punctuated pattern supporting an endocytotic internalization pathway as previously suggested.

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