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Index 399 Diffraction, 168 Disulphide bond formation, 126 DnaJ proteins, 305 DnaK/Hsp70 proteins, 305 Doxorubicin blood–brain barrier delivery quantification, 273–274 penetratins and, 38–40, 46 Dye exclusion permeability assays, 248–249 E 18A peptide, 202 Electron paramagnetic resonance (EPR) spectroscopy, 177, 233 Electrostatic potential, 194–195 Electrostatics, interfacial, 229–231 Endoplasmic reticulum (ER), targeting to, 303–304 Enzymatic viability assays, 251 Enzyme activity assays, quantification by, 270 Epidermal growth factor receptor (EGFR), quantification of bioactivity, 272–274 Escherichia coli SecA protein of, 299 Tat proteins in, 302 Eubacterial signal peptides, specificity, 308–309 Eukaryotic secretion pathways, 303–305 cotranslational, 303–304 targeting to ER, 303–304 translocon complex, 304 post-translational, 304–305 chaperone proteins, 305 membrane proteins in addition to Sec, 304–305 Eukaryotic signal peptides, specificity, 309–310 Experimental methods antimicrobial peptides, 385–388 antimicrobial activity assays, 385–386 cell uptake assays, 386–388 cell permeabilization assays, 386–388 fluorescence microscopy and FACS, 386 MAPs, 87–90 cell culture, 87 confocal laser scanning microscopy, 88–90 HPLC analysis, 88 uptake experiments, 87–88 penetratins, 40–47 AntpHD and penetratin, 40–42 AntpHD and penetratin-coupled cargoes, 42–46 comments, 46–47 F transportans, 66–68 cargo–CPP conjugation systems, 66 conjugation to proteins, 66–67 disulfide heterodimers, 66 Factor-independent bacterial protein secretion pathways, 302–303 energetic aspects, 302–303 insertion of procoat proteins, 302 Fhh and 4.55 RNA-dependent bacterial protein secretion pathways, 300–301 Flow cytometry, quantification by, 268 Fluorescein radiolabeling, 281 Fluorescein retention assay, 251 Fluorescence-activated cell sorting (FACS), 268 in microbial membrane-permeating peptides, 386 Fluorescence resonance energy transfer (FRET), 239 Fluorescence studies, 231, 238–239 of membrane interactions, 169–177 electron paramagnetic resonance (EPR) spectroscopy, 177 multidisciplinary monolayer-based approach, 177–177 in microbial membrane-permeating peptides, 386 quantification by, 267–269 Fourier transform infrared spectroscopy, 167–168, 232 4.5S RNA, 300 FtsY-dependent bacterial protein secretion pathways, 301 G Galanin, 54 Galparan, 53–55, see also Transportan Giant unicellular vesicles (GUVs), 227–228 G-proteins transportan and, 66–67 transportans and, 58 Grp170–Orp150 protein, 305 GTPases, small, 370 H Haemophilus influenzae, 299 Herpes simplex virus VP22 protein, 116 HIV Tat peptide, 127, 131, see also Tat entries Homeoprotein-derived peptidic vectors, penetratins and, 24–26 antennapedia homeodomain, 24–25
400 Cell-Penetrating Peptides: Processes and Applications AntpHD mutant behavior, 25 penetratin-1 peptide, 25–26 HPLC, quantification by, 269–270 3 H radiolabeling, quantification by, 266–267 h-region peptides, 117–122, see also Membrane translocating (MTS) peptides HSP70 proteins, 305 Hydrophobicity atomic surface, 191–193 molecular hydrophobicity potential (MHP), 199–202, 206–208 Hydrophobic membrane translocating sequence peptides, 115–140, see also Membrane translocating sequence (MTS) peptides Hydrophobic peptides, structure prediction modeling, 202–204 I 125 I radiolabeling, quantification by, 266–267 Immunodetection, quantification by, 270 Incubation, of cells with CPPs, 279–280 Inhibition zone assay, 385 Integrin β 3 h-region, 122 Interfacial electrostatics, 229–231 Internalization, 287–293, see also Uptake; Uptake kinetics comparisons of CPPs and, 290–291 penetratins, 26–30 blood–brain barrier and, 39–40 direct perfusion in CNS, 39 immune system, 39 in vivo with penatratin-derived vectors, 39–40 peptide–lipid interactions, 28–29 peptides in blood vessels, 39 proposed models, 29–30 structural parameters and translocation, 26–28 vesicular, 177 via receptor, 177–179 Interphase region, 229–231 Interpretative criteria, 239–240 Ion pump uptake assays, 252 K KALA peptide, 356–357 Kaposi fibroblast growth factor (kFGF, FGF-4), 117–118, 120–122, 131 Kar2p (BiP) protein, 305 KFFKFFKFFK peptide, 357 Kinetics, uptake, 277–293, see also Uptake kinetics L Lactate dehydrogenase leakage assay, 249 Large unicellular vesicles (LUVs), 227 Leakage, as mechanism of antimicrobial action, 383–384 Leakage assays, 283 Lhs1p protein, 305 Lipid perturbation, 193–194 Lipid vesicles, determination of reduction rate constants, 271–272 Liposomes, 226–228 Loligomers, in nucleic acid delivery, 357–358 Lymphocytes, transduction into, 370 M Magainin, 236–238, 240 Magnetic cell labeling with Tat protein, 336–343 CLIO-Tat internalization into lymphocyte and CD34+ subsets, 336–337 internalization of paramagnetic chelates, 342–343 internalization of superparamagnetic nanoparticles, 336 label distribution in dividing cell populations, 337–338 results in vitro, 338–339 results in vivo, 339–340 toxicity/nontoxicity, 338 in vivo MR imaging of Tat-labeled cells, 340–342 Maltoporin, 204–206 MAP and MAP analogues, uptake kinetics, 290 MAPs, 71–92 experimental methods, 87–90 cell culture, 87 confocal laser scanning microscopy, 88–90 HPLC analysis, 88 uptake experiments, 87–88 introducing history, 72–73 toxicity, 252–253 uptake mammalian cell experiments, 84–87 mechanistic aspects, 73–76 structural requirements, 76–84 Mastoparan, 58, 59, see also Transportan Mastoparan X, 236–238, 240 Mechanisms and interactions, see also specific topics backbone and side chain variations, 141–160 biophysical studies, 223–244 membrane interactions, 163–183 quantification of CPPs and cargoes, 263–275 signal peptides, 295–324
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CELL- PENETRATING PEPTIDES Processe
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Pharmacology and Toxicology: Basic
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Library of Congress Cataloging-in-P
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to the handbook are prominent resea
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REFERENCES 1. Green, M. and Loewens
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Contributors Mats Andersson Microbi
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Erin T. Pelkey Department of Chemis
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Contents Section I Classes of Cell-
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Chapter 16 Cell-Penetrating Peptide
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The Tat-Derived Cell-Penetrating Pe
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24 Cell-Penetrating Peptides: Proce
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3 Transportans Margus Pooga, Mattia
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Transportans 55 Indeed, galparan is
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Transportans 57 especially the endo
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Transportans 59 In the penetration
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Transportans 61 A 21-mer antisense
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Transportans 63 Commonly, the prote
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Transportans 65 antibiotin antibodi
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Transportans 67 For cross-linking o
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Transportans 69 and still retain ef
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Model Amphipathic Peptides 73 its D
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Model Amphipathic Peptides 75 pmol/
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TABLE 4.1 Internalization of Peptid
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TABLE 4.2 Internalization of Peptid
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Model Amphipathic Peptides 81 relat
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Model Amphipathic Peptides 87 A cat
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Model Amphipathic Peptides 89 At th
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Model Amphipathic Peptides 91 16. H
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Signal Sequence-Based Cell-Penetrat
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116 Cell-Penetrating Peptides: Proc
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Interactions of Cell-Penetrating Pe
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Structure Prediction of CPPs and It
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FIGURE 9.7 (Color Figure 9.7 follow
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FIGURE 9.8 The center of the figure
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Biophysical Studies of Cell-Penetra
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Toxicity and Side Effects of Cell-P
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Kinetics of Uptake of Cell-Penetrat
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Cell-Penetrating Peptide Conjugatio
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Page 370 and 371: Cell-Penetrating Peptides as Vector
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Page 400 and 401: Microbial Membrane-Permeating Pepti
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