PMID- 23919090
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20130806
LR  - 20211021
IS  - 1874-091X (Print)
IS  - 1874-091X (Electronic)
IS  - 1874-091X (Linking)
VI  - 7
DP  - 2013
TI  - Effects of redox state on the efficient uptake of cell permeable Peptide in 
      Mammalian cells.
PG  - 54-65
LID - 10.2174/1874091X20130531001 [doi]
AB  - We investigated whether a cell-penetrating peptide linked via a disulfide bond to 
      a fluorophore-labeled cargo peptide can be used to interrogate changes in 
      cellular redox state. A fluorescence resonance energy transfer (FRET) pair was 
      constructed so that the cargo peptide was labeled with fluorescein amidite (FAM) 
      and the cell-penetrating peptide was attached to a quencher. Incubation of cells 
      in culture with the FRET construct was visualized using live-cell, time-lapse 
      imaging, which demonstrated earlier cellular uptake of the construct when cells 
      were treated with the reducing agent n-acetylcysteine (NAC). The FRET peptide 
      construct was easily detected in cells cultured in 96-well plates using a 
      plate-reader. Treatment of cells with various classes of reducing or oxidizing 
      agents resulted in an increase or decrease in FAM fluorescence, respectively. 
      Changes in FAM fluorescence correlated significantly with redox-sensitive green 
      fluorescent protein ratios in cells treated with hydrogen peroxide but not NAC. 
      Detection of relative changes in cellular redox state was enhanced by the fact 
      that uptake of the cell-penetrating peptide occurred more quickly in relatively 
      reduced compared with oxidized cells. We conclude that cell-penetrating peptides 
      coupled via disulfide bonds to detectable cargo is a novel and specific approach 
      for assessment of relative changes in cellular thiol redox state.
FAU - Squires, Shayne
AU  - Squires S
AD  - Division of Cardiology, University of Utah School of Medicine, Salt Lake City, 
      Utah, USA.
FAU - Christians, Elisabeth
AU  - Christians E
FAU - Riedel, Michael
AU  - Riedel M
FAU - Timothy, David
AU  - Timothy D
FAU - Rodesch, Christopher K
AU  - Rodesch CK
FAU - Marvin, James
AU  - Marvin J
FAU - Benjamin, Ivor
AU  - Benjamin I
LA  - eng
GR  - DP1 OD006438/OD/NIH HHS/United States
GR  - R01 HL074370/HL/NHLBI NIH HHS/United States
GR  - T32 HL007576/HL/NHLBI NIH HHS/United States
PT  - Journal Article
DEP - 20130712
PL  - United Arab Emirates
TA  - Open Biochem J
JT  - The open biochemistry journal
JID - 101471222
PMC - PMC3731798
OTO - NOTNLM
OT  - FRET.
OT  - Redox
OT  - cell-penetrating peptides
OT  - glutathione
OT  - model amphipathic peptide
EDAT- 2013/08/07 06:00
MHDA- 2013/08/07 06:01
PMCR- 2013/01/01
CRDT- 2013/08/07 06:00
PHST- 2013/04/30 00:00 [received]
PHST- 2013/05/21 00:00 [revised]
PHST- 2013/05/27 00:00 [accepted]
PHST- 2013/08/07 06:00 [entrez]
PHST- 2013/08/07 06:00 [pubmed]
PHST- 2013/08/07 06:01 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - TOBIOCJ-7-54 [pii]
AID - 10.2174/1874091X20130531001 [doi]
PST - epublish
SO  - Open Biochem J. 2013 Jul 12;7:54-65. doi: 10.2174/1874091X20130531001. Print 
      2013.