crc press - E-Lib FK UWKS

Index 405
lipid perturbation (second restraint),
193–194
molecular hydrophobicity potential
(MHP), 199–202
Monte Carlo procedure, 198
Pex2Dstat files, 198–199
procedure, 197–198
principles and background, 188–190
results, 202–215
charged bilayer model, 210–215
efficiency of Monte Carlo method,
202–206
hydrophobic peptides, 202–204
membrane proteins, 204–206
penetratin, 206–207
uncharged membrane model, 207–210
Surface potential, 230
T
Targeting domains, for translocating peptides,
124–127
Tat
in nucleic acid delivery, 351–354
side effects, 256
Toxicity, 253
Tat-β-galactosidase, 372
Tat-Cre recombinase, 370–371
Tat-dependent bacterial protein secretion
pathways, 301–302
Tat-derived CPPs, 3–21
applications, 7–13
examples of vectorization, 17
principles and background, 3–4
uptake investigations and methods, 4–7
uptake mechanism, 13–16
cellular aspects, 16
of full-length Tat protein, 13
molecular aspects, 14–15
short HIV-Tat peptide, 13–14
Tat-E1A fusion protein, transduction into
lymphocytes, 370
Tat fusion proteins, cloning and purification,
368–369
Tat-GFP protein, 372
Tat protein, magnetic cell labeling with, 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
Tat PTD fusion zymogen, 372–373
Tat/Tat analogues, uptake kinetics, 289–290
Thermus aquaticus, 300–301
Thiazolidine ring formation protocol, 126–127
Toxicity, 179–180, 245–261, see also Side effects
in vitro findings, 252–255
arginine-rich peptides, 254
conclusions, 254
model amphipathic peptides (MAPs),
252–253
penetratin, 253–254
Tat, 253
transportan, 254
in vitro methods, 246–252
cell membrane permeability, 247–250
cytoplasmic leakage assays, 249–250
dye exclusion techniques, 248–249
cell viability assays, 250–252
enzymatic, 251
ion pump, 252
uptake, 251–252
in vivo, 256–258
Translocating chain-associated membrane
(TRAM) protein, 304
Translocation, possible mechanisms of, 240–241
Translocons, 297, 304
Transportan, 53–70, 128, 224–225
cell penetration, 55–57
discovery, 53–55
experimental methods, 66–68
cargo-CPP conjugation systems, 66
conjugation to proteins, 66–67
disulfide heterodimers, 66
125 I-radiolabeling, quantification by, 267
membrane-associated fraction, 282–283
in nucleic acid delivery, 354
properties, 57–58
sequences and uptake parameters, 54
side effects, 255–256
structural organization, 67–69
structure–activity relationships, 58–59
toxicity, 254
uptake kinetics, 285–287
as vector, 59–66
for peptides, 59–60
for PNA, 60–61
for proteins, 61–66
Transportan analogues, uptake kinetics, 287–289
Transverse relaxation optimized (TROSY)
spectroscopy, 235
Trojan horse strategies, 372–373