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398 Cell-Penetrating Peptides: Processes and Applications
liposomes and vesicles, 226–228
micelles, bicelles, and solvent mixtures,
228–229
interfacial phenomena — electrostatics,
229–231
interpretative criteria, 239–240
membrane effect comparison of CPPs and
other peptides, 236–238
methods, 231–233
circular dichroism, 232
electron spin (paramagnetic) resonance
(EPR) spectroscopy, 233
fluorescence, 231
Fourier transform infrared spectroscopy,
232
nuclear magnetic resonance (NMR),
232–233
possible mechanisms of translocation, 240–241
secondary structure induction by membranemimetic
solvents, 233–236
bicelles, 235
micelles, 234–235
positioning, 234–235
structure induction, 234
vesicles, 235–236
sequences and general properties of selected
CPPs, 223–225
translocation in model systems: penetratin,
238–239
Biotin/Abz/FITC-labeling, 268–269
Biotinylation and cell-ELISA, quantification by,
267
Blobel, Gunter, 296
Blood–brain barrier
quantification of bioactivity across, 273–274
Tat-β-galactosidase and, 372
Buforin, 236–238, 240
C
Cargo linkage, to translocating peptides, 124–127
Cationic translocating peptides, 127–129
CDC42 GTPases, 370
Cell-ELISA, quantification by, 267
Cell membrane permeability assays, 247–250
cytoplasmic leakage assays, 249–250
dye exclusion techniques, 248–249
Cell penetration, of transportans, 55–57
Cellular uptake, see Uptake
Cell viability assays, 250–252
enzymatic, 251
ion pump, 252
uptake, 251–252
Chaperone proteins, 305
Charged bilayer model, 210–215
Charge simulation, 194–197
Circular dichroism, 167, 232
Classes, see also specific classes
hydrophobic membrane translocating sequence
(MTS) peptides, 115–140
model amphipathic peptides (MAPs), 71–92
penetratins, 23–51
signal sequence-based CPPs and gene delivery,
93–113
Tat-derived CPPs, 3–21
transportans, 53–70
Confocal laser scanning microscopy, 72–73,
88–90, 281
Conjugation tactics, 328–336, see also Magnetic
cell labels
direct synthesis of CPP conjugates, 328–332
example: Tat-macrocytic chelator
conjugate, 329–332
magnetic cell labels and 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
solution phase conjugation, 332–336
amine-reactive reagents, 332–333
example: superparamagnetic iron oxide
particle–Tat conjugate, 334–336
heterobifunctional conjugation, 333–334
sulfhydryl-reactive reagents, 333
CS proteins, 130
Cytoplasmic leakage assays, 249–250
D
Debye length, 230
Delivery, penetratins, 31–40
AntpHD internalization of polypeptides, 34–35
chemical drug, 38
of entire proteins, 38
of peptide nucleic acids (PNAs), 37–38
principles of cargo–vector linkage, 31
vectorization
with AntpHD, 31–34
with penetratin peptides in vitro, 35–37
2-Deoxyglucose-6-phosphate (DGP) leakage
assay, 250