Quantitative evaluation of the cell penetrating properties of an iodinated Tyr-L-maurocalcine analog
| Autor: | Pascale Perret, Michel Ronjat, Lucie Dardevet, Michel De Waard, Catherine Ghezzi, Yves Usson, Mitra Ahmadi, Daniel Fagret, Sandrine Bacot, Céline Tisseyre |
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| Přispěvatelé: | [GIN] Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Joseph Fourier - Grenoble 1 (UJF), Radiopharmaceutiques biocliniques (LRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Laboratoire de Physique Corpusculaire - Clermont-Ferrand (LPC), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Dynamique Cellulaire et Tissulaire- Interdisciplinarité, Modèles & Microscopies (TIMC-IMAG-DyCTiM), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Dynamiques Cellulaire, Tissulaire & Microscopie fonctionnelle (TIMC-IMAG-DyCTiM), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular Protein Folding Cell Membrane Permeability [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging Cell Cell-Penetrating Peptides MESH: Amino Acid Sequence Radioiodination MESH: Tyrosine Iodine Radioisotopes Cytosol 0302 clinical medicine MESH: Scorpion Venoms MESH: Cytosol MESH: Animals MESH: Cell Membrane Permeability MESH: Cell Size MESH: Cell-Penetrating Peptides Drug Carriers 0303 health sciences MESH: Kinetics Chemistry MESH: Iodine Radioisotopes 3. Good health MESH: Drug Carriers medicine.anatomical_structure Maurocalcine Biochemistry 030220 oncology & carcinogenesis Cell fractionation Intracellular MESH: Models Molecular MESH: Cell Nucleus Quantitative evaluation MESH: Solid-Phase Synthesis Techniques MESH: Cell Line Tumor MESH: Rats MESH: Biological Transport MESH: Protein Folding Molecular Sequence Data Scorpion Venoms 03 medical and health sciences Cell Line Tumor medicine Extracellular Animals Amino Acid Sequence Molecular Biology Solid-Phase Synthesis Techniques Cell Size 030304 developmental biology Cell Nucleus Cell penetrating peptide MESH: Molecular Sequence Data Cell Membrane Biological Transport Cell Biology Rats Kinetics Cytoplasm Drug delivery Cell-penetrating peptide Tyrosine MESH: Cell Membrane |
| Zdroj: | BBA-Biochimica et Biophysica Acta BBA-Biochimica et Biophysica Acta, Elsevier, 2014, 1843 (10), pp.2356-64. ⟨10.1016/j.bbamcr.2014.03.017⟩ BBA-Biochimica et Biophysica Acta, 2014, 1843 (10), pp.2356-64. ⟨10.1016/j.bbamcr.2014.03.017⟩ |
| ISSN: | 0006-3002 |
| DOI: | 10.1016/j.bbamcr.2014.03.017⟩ |
| Popis: | International audience; L-Maurocalcine (L-MCa) is the first reported animal cell-penetrating toxin. Characterizing its cell penetration properties is crucial considering its potential as a vector for the intracellular delivery of drugs. Radiolabeling is a sensitive and quantitative method to follow the cell accumulation of a molecule of interest. An L-MCa analog containing an additional N-terminal tyrosine residue (Tyr-L-MCa) was synthesized, shown to fold and oxidize properly, and successfully radioiodinated to (125)I-Tyr-L-MCa. Using various microscopy techniques, the average volume of the rat line F98 glioma cells was evaluated at 8.9 to 18.9×10(-7)μl. (125)I-Tyr-L-MCa accumulates within cells with a dose-dependency similar to the one previously published using 5,6-carboxyfluorescein-L-MCa. According to subcellular fractionation of F98 cells, plasma membranes keep less than 3% of the peptide, regardless of the extracellular concentration, while the nucleus accumulates over 75% and the cytosol around 20% of the radioactive material. Taking into account both nuclear and cytosolic fractions, cells accumulate intracellular concentrations of the peptide that are equal to the extracellular concentrations. Estimation of (125)I-Tyr-L-MCa cell entry kinetics indicate a first rapid phase with a 5min time constant for the plasma membrane followed by slower processes for the cytoplasm and the nucleus. Once inside cells, the labeled material no longer escapes from the intracellular environment since 90% of the radioactivity remains 24h after washout. Dead cells were found to have a lower uptake than live ones. The quantitative information gained herein will be useful for better framing the use of L-MCa in biotechnological applications. This article is part of a Special Issue entitled: Calcium Signaling in Health and Disease. Guest Editors: Geert Bultynck, Jacques Haiech, Claus W. Heizmann, Joachim Krebs, and Marc Moreau. |
| Databáze: | OpenAIRE |
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