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406 Cell-Penetrating Peptides: Processes and Applications
Trypsinization, of surface CPP, 280
Tryptophan, fluorescence studies, 231
U
Uncharged membrane model, 207–210
Unknown or factor-independent bacterial protein
secretion pathways, 302–303
energetic aspects, 302–303
insertion of procoat proteins, 302
Uptake
backbone and side chain variations and,
141–160, see also Backbone and
side chain variations
discussion, 156–159
materials and methods, 144–147
principles and background, 141–144
results, 147–156
MAPs
mammalian cell experiments, 84–87
mechanistic aspects, 73–76
structural requirements, 76–84
Tat-derived CPPs
investigations and methods, 4–7
mechanism, 13–16
cellular aspects, 16
of full-length Tat protein, 13
molecular aspects, 14–15
short HIV-Tat peptide, 13–14
Uptake assays, 251–252
Uptake kinetics, 277–293
conclusions, 291
experimental results, 285–291
comparative internalization,
290–291
MAP and MAP analogues, 290
penetratin/penetratin analogues, 289
Tat/Tat analogues, 289–290
transportan, 285–287
transportan analogues, 287–289
methodological considerations, 278–283
cell incubation with CPP, 278–280
cells, 278
detection of CPP, 280–282
exclusion of pore formation, 283
V
internalization data, 283
quantification in cell membranes, 282–283
principles and background, 277–278
types of experiments and data analysis,
283–285
progress curves at different CPP
concentrations, 285
single time point measurements, 283–284
uptake measurements as function of time,
284–285
Vectorization
of Tat-derived CPPs, 17
of transportan, 59–66
for peptides, 59–60
for PNA, 60–61
for proteins, 61–66
Vesicles, 226–228, 235–236
giant unicellular (GUVs), 227–228
large unicellular (LUVs), 227
lipid, determination of reduction rate constants,
271–272
small unicellular (SUVs), 227
Vesicular internalization, 177
VP22 protein, 116
W
Water–bilayer interface description, 190–191
Weight matrix prediction methods, 311
X
XcpA (PilD) signal peptidase, 307
Y
Ydji1p protein, 305
YidC protein, 300, 302
Z
Zymogen, Tat PTD fusion, 372–373