細胞穿透胜肽之分子運送
Molecular Delivery of CellPenetrating Peptides
李漢榮
國立東華大學自然資源與環境學系
Speaker: Dr. Han-Jung Lee
Department of Natural Resources and
Environmental Studies
National Dong Hwa University
1
Introduction
b-Gal
PTD
Extracellular
Intracellular
Plasma
Membrane
Schwarze et al., Science 285: 1569-1572, 1999
2
Cell-Penetrating Peptides
(CPPs)
Transactivator (Tat) protein of the
human immunodeficiency virus type 1
(HIV-1) was able to enter into cells and
activate HIV-LTR.
Green and Loewenstein, Cell 55: 1179-1188, 1988
Frankel and Pabo, Cell 55: 1189-1193, 1988
3
Cell-Penetrating Peptides
(CPPs)
DNA
RNA
Tat
O2
H2O
protein
Extracellular
Intracellular
Plasma
Membrane
4
Cell-Penetrating Peptides
(CPPs)
Functional domains of HIV-1 Tat :
CxxCxCxxCCxxCxxC
1
2
11
22
acidic
37
cysteinerich
47
core
57
72
86
basic augmenting
EPVDPRLEPW
YGRKKRRQRRR
Basic amino acid-rich domain
Gaynor, Curr. Top. Microbiol. Immunol. 193: 51-77, 1995
Vives et al., J. Biol. Chem. 272: 16010-16017, 1997
Cell-Penetrating Peptides
(CPPs)
CPPs, also called protein transduction
domains (PTDs), are a group of short, highly
basic peptides which can penetrate into cells
either alone or associated with cargoes.
Tat
YGRKKRRQRRR
(CPP)
6
Results
Chang et al., Plant Cell Physiol. 46: 482-488, 2005
7
GFP
(200x)
Tat-GFP
R9-GFP
(200x)
(200x)
Tat-GFP
R9-GFP
(400x)
(400x)
Covalent protein transduction into onion root tip cells
8
RFP
(onion epidermal cells)
R9-RFP
(tomato root cells)
R9-RFP
(onion epidermal
cells)
Covalent protein transduction in plant cells
9
Covalent and Noncovalent Protein
Transductions
protein
covalent protein
transduction
(CPT)
protein
noncovalent protein
transduction
(NPT)
Plasma Membrane
10
GFP
R9/GFP
RFP
R9/RFP
Noncovalent protein transduction in onion root cells
Chang et al., New Phytol. 174: 46-56, 2007
11
Cell-Penetrating Peptides
(CPPs)
CPPs are able to deliver proteins into
cells in a covalent protein
transduction (CPT) and noncovalent
protein transduction (NPT).
12
Cell-Penetrating Peptides
(CPPs)


Synthetic nona-arginine (SR9)
N-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-C
13
SR9
(mung bean)
SR9/DNA
(mung bean)
DNA
(soybean)
DNA
(mung bean)
SR9/DNA
(mung bean)
SR9/DNA
(soybean)
Chen et al., FEBS Lett. 581: 1891-1897, 2007
14
dsRNA
GC-EGFP cells
SR9/
dsRNA
Wang et al., J. Biochem. Biophy. Methods 70: 579-586, 2007
15
Cell-Penetrating Peptides
(CPPs)
CPPs can deliver proteins, DNAs and
RNAs into cells in a noncovalent
manner.
16
GFP
RFP
PBS
R9-GFP
R9-GFP/RFP
Hu et al., Peptides 30: 1669-1678, 2009
RFP
R9-GFP
R9-GFP/RFP
17
Cell-Penetrating Peptides
(CPPs)
CPPs are able to deliver proteins into
cells in a covalent protein
transduction (CPT), noncovalent
protein transduction (NPT) and
covalent and noncovalent protein
transductions (CNPT).
18
Quantum
dots
Lyso
Tracker
Hoechst
(merge)
Liu et al., Biomaterials 32: 3520-3537, 2011
19
Summary
We demonstrate that CPPs can
effectively deliver proteins, DNAs,
RNAs and nanoparticles into live
cells.
17
20
Summary
However, the mechanism of protein
transduction is unknown yet.
21
CPP Translocation Mechanisms
Cargo
CPP