Noncovalent protein transduction in plant cells by macropinocytosis

Chang, Mei‐Chi; Chou, Jyh-Ching; Chen, Chung‐Pin; Liu, Betty Revon; Lee, Han-Jung

doi:10.1111/j.1469-8137.2007.01977.x

Cited by 74 publications

(112 citation statements)

References 34 publications

(60 reference statements)

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“…We did not find any significant effect of macropinocytosis inhibitors on the uptake of pVEC and transportan in mesophyll protoplasts (Figure 3(B)). However, noncovalent protein transduction of fluorescent proteins using arginine-rich CPPs in plant root tip cells is inhibited in the presence of the same macropinocytic inhibitors [13].…”

Section: Resultsmentioning

confidence: 99%

“…[11]. Fluorescent proteins have been delivered in plant cells either by fusion or noncovalent interaction with arginine-rich intracellular delivery (AID) peptides [12,13]. We have also shown that monomer and dimer of HIV-1 Tat basic domain translocate across plasma membrane and accumulate in the nuclei of triticale mesophyll protoplasts [14].…”

Section: Introductionmentioning

confidence: 99%

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Cellular uptake of cell‐penetrating peptides pVEC and transportan in plants

Chugh

Eudes

2007

Journal of Peptide Science

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Internalization of fluorescently labeled CPPs, pVEC, transportan and scrambled pVEC, in a range of plant cells was investigated. Cellular uptake of the peptides was found to be tissue dependent. pVEC and transportan were distinctly internalized in triticale mesophyll protoplasts, onion epidermal cells, leaf bases and root tips of seven-day old triticale seedlings but showed negligible florescence in coleoptile and leaf tips as observed under a fluorescence microscope. Further, pVEC and transportan uptake studies were focused on mesophyll protoplasts as a system of investigation. In fluorimetric studies transportan showed 2.3 times higher cellular internalization than pVEC in protoplasts, whereas scrambled pVEC failed to show any significant fluorescence. Effect of various factors on cellular internalization of pVEC and transportan in protoplasts was also investigated. The cellular uptake of both the peptides was concentration dependent and nonsaturable. The cellular uptake of pVEC and transportan was enhanced at low temperature (4°C). The presence of endocytic/macropinocytosis inhibitors did not reduce the cellular uptake of the peptides, suggesting direct cell penetration, receptor-independent internalization of pVEC and transportan into the plant cells.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cellular uptake of cell‐penetrating peptides pVEC and transportan in plants

Chugh

Eudes

2007

Journal of Peptide Science

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“…Characterization of the mechanisms by which PTDs transport proteins suggests that an electrostatic A, avirulent; V, virulent; NT, not tested; Y, significantly fewer blue (GUS-positive) tissue patches from GUS expression resulting from Avr1b-induced cell death; N, not significantly fewer blue tissue patches; P, partial reduction in blue tissue patches; SP, signal peptide. interaction between cationic PTDs and the anionic surface of the plasma membrane leads to transport via a specialized form of endocytosis called macropinocytosis (Snyder and Dowdy, 2004;Kaplan et al, 2005) that occurs in plant cells (Chang et al, 2007) and animal cells. Thus, macropinocytosis is one candidate mechanism by which RXLR-dEER proteins might enter cells.…”

Section: Discussionmentioning

confidence: 99%

“…If mutations were present in the RXLR or dEER motifs of the fusion protein, GFP did not accumulate inside the soybean root cells (Figures 4E and 4F,respectively). When the RXLR-dEER region was replaced by the artificial protein transduction motif Arg 9 (Chang et al, 2005(Chang et al, , 2007; see below), GFP once again entered the soybean root cells ( Figure 4E) and accumulated in the nuclei ( Figure 4H). …”

Section: Rxlr-mediated Transit Into Soybean Cells Does Not Require Thmentioning

confidence: 99%

RXLR-Mediated Entry of Phytophthora sojae Effector Avr1b into Soybean Cells Does Not Require Pathogen-Encoded Machinery

et al. 2008

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Effector proteins secreted by oomycete and fungal pathogens have been inferred to enter host cells, where they interact with host resistance gene products. Using the effector protein Avr1b of Phytophthora sojae, an oomycete pathogen of soybean (Glycine max), we show that a pair of sequence motifs, RXLR and dEER, plus surrounding sequences, are both necessary and sufficient to deliver the protein into plant cells. Particle bombardment experiments demonstrate that these motifs function in the absence of the pathogen, indicating that no additional pathogen-encoded machinery is required for effector protein entry into host cells. Furthermore, fusion of the Avr1b RXLR-dEER domain to green fluorescent protein (GFP) allows GFP to enter soybean root cells autonomously. The conclusion that RXLR and dEER serve to transduce oomycete effectors into host cells indicates that the >370 RXLR-dEER–containing proteins encoded in the genome sequence of P. sojae are candidate effectors. We further show that the RXLR and dEER motifs can be replaced by the closely related erythrocyte targeting signals found in effector proteins of Plasmodium, the protozoan that causes malaria in humans. Mutational analysis of the RXLR motif shows that the required residues are very similar in the motifs of Plasmodium and Phytophthora. Thus, the machinery of the hosts (soybean and human) targeted by the effectors may be very ancient.

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“…Covalent and noncovalent transduction of small 24 kDa fluorescent reporter proteins by Tat-PTD and arginine-rich intracellular delivery (AID) proteins in mung bean, soybean, corn, and onion root tip cells were reported by Chang et al (8,101). The cell wall remains a major obstacle for transduction of larger proteins in plant tissues, such as immature scutellum and cotyledon, a model system for genetic transformation studies owing to their amenability toward tissue culture procedures and high efficacy for plant regeneration.…”

Section: Somatic Cellsmentioning

confidence: 99%

Cell‐penetrating peptides: Nanocarrier for macromolecule delivery in living cells

2010

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SummaryNovel classes and applications of cell-penetrating peptides (CPPs) are being constantly discovered since they were first identified 2 decades ago. These short cationic peptides (nanomolecules) either by covalent binding or by noncovalent binding can traverse cell membranes and deliver a variety of molecules that are unable to overcome the permeability barrier in their own capacity. The ability of the CPPs to deliver variety of macromolecules, such as oligonucleotides, therapeutic drugs, proteins, and medical imaging agents, by forming nanoparticulate carriers in a range of cells has led them to emerge as a potential tool for both macromolecule delivery application and to gain insight into the fundamentals of mechanism of cellular uptake across the plasma membrane. This review explores the recent advances, challenges, and future prospects in the field of CPP-mediated cargo delivery in mammalian and plant cells. Studies have been conducted into the peptide chemistry and stability of CPP-macromolecular complexes. Most of the CPPs have been shown to be nontoxic and do not interfere with the functionality of the macromolecules delivered across the cell membrane. The mechanism of uptake of CPP-cargo complexes and the uptake of CPPs alone across the plasma membrane remains unresolved. As the world of CPPs is rapidly advancing in both mammalian and plant system, there is a promising future for the various applications of transduction and transfection into intact cells.

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Noncovalent protein transduction in plant cells by macropinocytosis

Cited by 74 publications

References 34 publications

Cellular uptake of cell‐penetrating peptides pVEC and transportan in plants

Cellular uptake of cell‐penetrating peptides pVEC and transportan in plants

RXLR-Mediated Entry of Phytophthora sojae Effector Avr1b into Soybean Cells Does Not Require Pathogen-Encoded Machinery

Cell‐penetrating peptides: Nanocarrier for macromolecule delivery in living cells

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