Molecular delivery of cell penetrating peptides
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細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides PowerPoint PPT Presentation


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細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating 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. b -Gal. Introduction. PTD. Extracellular. Intracellular. Plasma Membrane.

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細胞穿透胜肽之分子運送 Molecular Delivery of Cell-Penetrating Peptides

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Molecular delivery of cell penetrating peptides

細胞穿透胜肽之分子運送Molecular Delivery of Cell-Penetrating 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


Molecular delivery of cell penetrating peptides

b-Gal

Introduction

PTD

Extracellular

Intracellular

Plasma Membrane

Schwarze et al., Science 285: 1569-1572, 1999


Molecular delivery of cell penetrating peptides

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


Molecular delivery of cell penetrating peptides

DNA

RNA

protein

Tat

O2

H2O

Cell-Penetrating Peptides

(CPPs)

Extracellular

Intracellular

Plasma Membrane


Molecular delivery of cell penetrating peptides

Cell-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

CxxCxCxxCCxxCxxC

1

11

22

37

72

2

47

57

86

cysteine-rich

acidic

basic

core

augmenting

EPVDPRLEPW

YGRKKRRQRRR

Basic amino acid-rich domain


Molecular delivery of cell penetrating peptides

Tat

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.

YGRKKRRQRRR

(CPP)


Molecular delivery of cell penetrating peptides

Results

Chang et al., Plant Cell Physiol. 46: 482-488, 2005


Molecular delivery of cell penetrating peptides

GFP (200x)

R9-GFP (200x)

Tat-GFP (200x)

R9-GFP (400x)

Tat-GFP (400x)

Covalent protein transduction into onion root tip cells


Molecular delivery of cell penetrating peptides

R9-RFP

RFP

(tomato root cells)

(onion epidermal cells)

R9-RFP

(onion epidermal

cells)

Covalent protein transduction in plant cells


Covalent and noncovalent protein transductions

protein

protein

Covalent and Noncovalent Protein Transductions

covalent protein transduction (CPT)

noncovalent protein transduction (NPT)

Plasma Membrane


Molecular delivery of cell penetrating peptides

GFP

R9/GFP

RFP

R9/RFP

Noncovalent protein transduction in onion root cells

Chang et al., New Phytol. 174: 46-56, 2007


Molecular delivery of cell penetrating peptides

Cell-Penetrating Peptides

(CPPs)

CPPs are able to deliver proteins into cells in a covalent protein transduction (CPT) and noncovalent protein transduction (NPT).


Molecular delivery of cell penetrating peptides

Cell-Penetrating Peptides

(CPPs)

  • Synthetic nona-arginine (SR9)

  • N-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-C


Molecular delivery of cell penetrating peptides

DNA

SR9

(mung bean)

(mung bean)

SR9/DNA

SR9/DNA

(mung bean)

(mung bean)

DNA

SR9/DNA

(soybean)

(soybean)

Chen et al., FEBS Lett. 581: 1891-1897, 2007


Molecular delivery of cell penetrating peptides

GC-EGFP cells

dsRNA

SR9/ dsRNA

Wang et al., J. Biochem. Biophy. Methods 70: 579-586, 2007


Molecular delivery of cell penetrating peptides

Cell-Penetrating Peptides

(CPPs)

CPPs can deliver proteins, DNAs and RNAs into cells in a noncovalent manner.


Molecular delivery of cell penetrating peptides

GFP

RFP

PBS

RFP

R9-GFP

R9-GFP

R9-GFP/RFP

R9-GFP/RFP

Hu et al., Peptides 30: 1669-1678, 2009


Molecular delivery of cell penetrating peptides

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).


Molecular delivery of cell penetrating peptides

Quantum dots

Lyso Tracker

Hoechst

(merge)

Liu et al., Biomaterials 32: 3520-3537, 2011


Molecular delivery of cell penetrating peptides

Summary

We demonstrate that CPPs can effectively deliver proteins, DNAs, RNAs and nanoparticles into live cells.

17


Molecular delivery of cell penetrating peptides

Summary

However, the mechanism of protein transduction is unknown yet.


Cpp translocation mechanisms

CPP Translocation Mechanisms

Cargo CPP


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