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More than a decade ago, it was discovered that cationic peptides could traverse the cellular plasma membrane without specific transporter proteins or membrane damage. Subsequently, it was found that these peptides, known as cell-penetrating peptides (CPPs), were also capable of delivering cargos into cells, hence the great potential of these vectors was acknowledged. Today, many different research groups are working with CPPs, which necessitates efforts to develop unified assays enabling the comparison of data. Here we contribute three protocols for evaluation of CPPs which, if used in conjunction, provide complementary data about the amount and mechanism of uptake (fluorometric analysis and confocal microscopy, respectively), as well as the extent of degradation (HPLC analysis of cell lysates). All three protocols are based on the use of fluorescently labeled peptides and can be performed on the same workday.
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http://dx.doi.org/10.1038/nprot.2006.174 | DOI Listing |
Int J Biol Macromol
March 2025
College of Chemistry and Chemical Engineering, Yunnan Normal University, Kunming, Yunnan, China. Electronic address:
Liquid-liquid phase separation (LLPS) of nuclear pore complex (NPC) with nuclear transport proteins (NTPs) via intrinsically disordered regions (IDRs) plays a crucial role in the nucleocytoplasmic transport. The development of efficient targeted delivery systems based on LLPS has attracted widespread attention. Here, we developed nanocarriers of casein peptides, a natural intrinsically disordered proteins (IDPs), modified with fatty acids of different alkyl chains (C10-C18) and decorated by shellac for highly effective drug delivery and cancer therapy.
View Article and Find Full Text PDFAm J Respir Cell Mol Biol
March 2025
Indiana University Purdue University at Indianapolis, Indianapolis, Indiana, United States;
High Mobility Group Box 1 (HMGB1) is a nuclear protein released from damaged cells and implicated in the pathogenesis of Pulmonary Arterial Hypertension (PAH) through activation of pro-inflammatory and pro-survival responses. However, the role of intracellular HMGB1 signaling, particularly its interactions with DNA and transcriptional regulation, remains underexplored. In this study, we investigated the role of intracellular HMGB1 and its critical residue Cys106 by engineering cell-penetrating peptide (αHMGB1Cys106) that mimics part of the HMGB1 dimeric interface surrounding Cys106.
View Article and Find Full Text PDFStem Cell Res Ther
March 2025
Advanced Therapy Medicinal Product Technology Development Center (ATMP-TDC), Cell Science Research Center, ACECR, Royan Institute for Stem Cell Biology and Technology, Tehran, Iran.
Background: Diabetes mellitus remains a pervasive global health concern, urging a deeper exploration of islet transplantation as a potential enduring solution. The efficacy of this therapeutic approach pivots on the precision of cryopreservation techniques, ensuring both the viability and accessibility of pancreatic islets. This study delves into the merits of cryopreserving these islets using the disaccharide trehalose, accompanied by an inventive strategy involving poly L proline (PLP) as a cell-penetrating peptide to overcome the cryoprotectant limitations inherent to trehalose.
View Article and Find Full Text PDFBiochemistry
March 2025
Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Queensland, Brisbane, QLD 4072, Australia.
Peptides that have a head-to-tail cyclic backbone tend to be more stable than linear peptides, as do peptides that contain one or more cross-linking disulfide bond. Some of these cyclic and/or disulfide rich peptides have been reported to penetrate cells. These include peptides from a wide range of natural sources, including plants, spiders, crabs, and humans.
View Article and Find Full Text PDFInt J Mol Sci
February 2025
Department of Organic Chemistry, Institute of Chemistry, Faculty of Science, ELTE Eötvös Loránd University, Pázmány Péter Sétány 1/A, 1117 Budapest, Hungary.
Getting through the cell membrane is challenging, and transporting a therapeutic agent while entering the cell is even more complicated. Cell-penetrating peptides (CPPs) are valuable tools for solving this problem, although they have drawbacks. In this work, the synthesis and investigation of efficient CPPs are described.
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