Self-assembled complex of probe peptide--E. Coli RNA I conjugate and nano graphene oxide for apoptosis diagnosis

Biomaterials. 2012 Oct;33(30):7556-64. doi: 10.1016/j.biomaterials.2012.06.086. Epub 2012 Jul 20.

Abstract

Caspase-3 plays an important role in the initiation and propagation of apoptosis which is involved in various kinds of diseases including neurodegenerative diseases and inflammatory diseases. The capase-3 cleavable site of Asp-Glu-Val-Asp (DEVD) connected to a partial sequence of Escherichia coli RNAI was labeled with tetramethyl-6-carboxyrhodamine (TAMRA) as an optical probe. Graphene oxide (GO) was synthesized by a modified Hummer's method and exploited for the preparation of nano-sized GO (NGO) conjugated with polyethylene glycol (PEG). After binding the NGO-PEG by π-π stacking, the quenched fluorescence of TAMRA-DEVD-single stranded DNA (ssDNA) conjugate was recovered via the enzymatic cleavage by caspase-3 in live A549 cells. The comparative study with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay clearly confirmed the specific detection of apoptosis by the non-covalent TAMRA-DEVD-ssDNA/NGO-PEG complex. Furthermore, the self-assembled NGO complex was successfully exploited for in vivo diagnosis of apoptosis-related diseases like hypoxic-ischemic encephalopathy (HIE) and liver cirrhosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Brain / pathology
  • Brain Ischemia / diagnosis
  • Brain Ischemia / pathology
  • Caspase 3 / metabolism
  • Cell Line
  • Escherichia coli / genetics*
  • Graphite*
  • Humans
  • Hypoxia / diagnosis
  • Intracellular Space / enzymology
  • Kidney / pathology
  • Liver / pathology
  • Liver Cirrhosis / diagnosis
  • Liver Cirrhosis / pathology
  • Microscopy, Atomic Force
  • Microscopy, Confocal
  • Nanoparticles* / ultrastructure
  • Peptides*
  • Polyethylene Glycols / chemistry
  • RNA, Bacterial*
  • Rats
  • Rats, Sprague-Dawley
  • Spectrometry, Fluorescence

Substances

  • Peptides
  • RNA I
  • RNA, Bacterial
  • Polyethylene Glycols
  • Graphite
  • Caspase 3