細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides
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Transcript of 細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides
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細胞穿透胜肽之分子運送細胞穿透胜肽之分子運送Molecular Delivery of Cell-Molecular Delivery of Cell-
Penetrating PeptidesPenetrating Peptides
李漢榮李漢榮
國立東華大學國立東華大學自然資源與環境學系自然資源與環境學系
Speaker: Dr. Han-Jung LeeSpeaker: Dr. Han-Jung LeeDepartment of Natural Resources and Department of Natural Resources and
Environmental StudiesEnvironmental StudiesNational Dong Hwa UniversityNational Dong Hwa University
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Plasma Plasma MembraneMembrane
Extracellular Intracellular
IntroductionIntroduction
Schwarze et al., Science 285: 1569-1572, 1999
-Gal
PTD
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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
Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
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DNA
RNA
protein
Tat
Plasma Plasma MembraneMembrane
O2
H2O
Extracellular Intracellular
Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
Functional domains of HIV-1 Tat :
Gaynor, Curr. Top. Microbiol. Immunol. 193: 51-77, 1995
Vives et al., J. Biol. Chem. 272: 16010-16017, 1997
acidiccysteine-rich core basic augmenting
2 11 22 37 47 57 721
CxxCxCxxCCxxCxxC
YGRKKRRQRRREPVDPRLEPW
86
Basic amino acid-rich domain
Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
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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.
TatYGRKKRRQRRR
(CPP)
Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
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ResultsResults
Chang et al., Plant Cell Physiol. 46: 482-488, 2005
8Covalent protein transduction into onion root tip cells
Tat-GFP (400x)
R9-GFP (400x)
Tat-GFP (200x)
R9-GFP (200x)
GFP
(200x)
9Covalent protein transduction in plant cells
R9-RFP
R9-RFP
RFP(onion epidermal cells)
(onion epidermalcells)
(tomato root cells)
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protein
covalent protein transduction
(CPT)
noncovalent protein transduction
(NPT)
Covalent and Noncovalent Protein Covalent and Noncovalent Protein TransductionsTransductions
Plasma MembranePlasma Membrane
protein
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GFP
RFP
R9/GFP
R9/RFP
Noncovalent protein transduction in onion root cells
Chang et al., New Phytol. 174: 46-56, 2007
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Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
CPPs are able to deliver proteins into cells in a covalent protein transduction (CPT) and noncovalent protein transduction (NPT).
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Synthetic nona-arginine (SR9)Synthetic nona-arginine (SR9) N-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-CN-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-C
Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
14Chen et al., FEBS Lett. 581: 1891-1897, 2007
SR9 DNA
SR9/DNA SR9/DNA
SR9/DNADNA
(mung bean)
(mung bean)
(mung bean) (mung bean)
(soybean) (soybean)
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GC-EGFP cellsdsRNA
SR9/ dsRNA
Wang et al., J. Biochem. Biophy. Methods 70: 579-586, 2007
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Cell-Penetrating PeptidesCell-Penetrating Peptides(CPPs)
CPPs can deliver proteins, DNAs and RNAs into cells in a noncovalent manner.
17Hu et al., Peptides 30: 1669-1678, 2009
PBS
GFP RFP
RFP
R9-GFP R9-GFP
R9-GFP/RFP R9-GFP/RFP
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Cell-Penetrating PeptidesCell-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).
19Liu et al., Biomaterials 32: 3520-3537, 2011
Lyso Tracker
Hoechst
(merge)
Quantum dots
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Summary
We demonstrate that CPPs can effectively deliver proteins, DNAs, RNAs and nanoparticles into live cells. 17
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Summary
However, the mechanism of protein transduction is unknown yet.
CPP Translocation CPP Translocation MechanismsMechanisms
Cargo CPP