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Article 17
Enhanced biomolecular detection based on localized
surface plasmon resonance (LSPR) using
enzyme-precipitation reaction
Article 18
Microglial peroxiredoxin V acts as an inducible
anti-inflammatory antioxidant through cooperation
with redox signaling cascades
J Nanosci Nanotechnol. 2010 May; 10(5):3246-9.
J Neurochem. 2010 Jul; 114(1):39-50.
Lee SW, Ahn J, Kim MG, Shin YB * , Lee JJ, Lim KP, Kim
KB
* Correspondence: ybshin@kribb.re.kr
BioMonitoring Research Center
An enzyme-catalyzed precipitation reaction was employed
as a means to increase the change in the LSPR signal after
intermolecular bindings between antigens and antibodies occurred
on gold nanodot surfaces. The gold nanodot array
with an diameter of 175 nm and a thickness of 20 nm was
fabricated on a glass wafer using thermal nanoimprint
lithography. The human interleukin (hIL) 5 antibody was
immobilized on the gold nanodot, followed by binding of
hIL 5 to the anti-hIL 5. Subsequently, a biotinylated anti-hIL
5 and a alkaline phosphatase conjugated with streptavidin
were simultaneously introduced. A mixture of 5-bromo-4-chloro-3-indolyl
phosphate p-toluidine (BCIP) and nitro
blue tetrazolium (NBT) was then used for precipitation,
which resulted from the biocatalytic reaction of the alkaline
phosphatase on gold nanodot. The LSPR spectra were obtained
after each binding process. Using this analysis, the
enzyme-catalyzed precipitation reaction on gold nanodots
was found to be effective in amplifying the change in the
peak wavelength of LSPR after molecular bindings.
PMID:20358932
Keywords: Biopolymers; Biosensing techniques;
Enzyme-catalyzed precipitation; Equipment design;
Immunoassay; Metalloproteins; Nanodot;
Nanoimprint lithography (NIL); Nanotubes; Surface
plasmon resonance
Sun HN, Kim SU, Huang SM, Kim JM, Park YH, Kim
SH, Yang HY, Chung KJ, Lee TH, Choi HS, Min JS, Park
MK, Kim SK, Lee SR, Chang KT, Lee SH, Yu DY * , Lee
DS
* Correspondence: dyyu10@kribb.re.kr
Aging Research Center
Reactive oxygen species (ROS) actively participate in microglia-mediated
pathogenesis as pro-inflammatory molecules.
However, little is known about the involvement of specific
antioxidants in maintaining the microglial oxidative balance.
We demonstrate that microglial peroxiredoxin (Prx) 5 expression
is up-regulated by lipopolysaccharide (LPS) through
activation of the ROS-sensitive signaling pathway and is
involved in attenuation of both microglial activation and
nitric oxide (NO) generation. Unlike in stimulation of oxidative
insults with paraquat and hydrogen peroxide, Prx V
expression is highly sensitive to LPS-stimulation in
microglia. Reduction of ROS level by treatment with either
NADPH oxidase inhibitor or antioxidant ablates LPS-mediated
Prx V up-regulation in BV-2 microglial cells and is
closely associated with the activation of the c-jun N-terminal
kinase (JNK) signaling pathway. This suggests the involvement
of ROS/JNK signaling in LPS-mediated Prx V
induction. Furthermore, NO induces Prx V up-regulation
that is ablated by the addition of inducible nitric oxide synthase
inhibitor or deleted mutation of inducible nitric oxide
synthase in LPS-stimulated microglia. Therefore, these results
suggest that Prx V is induced by cooperative action
among the ROS, RNS, and JNK signaling cascades.
Interestingly, knockdown of Prx V expression causes the
acceleration of microglia activation, including augmented
ROS generation and JNK-dependent NO production. In summary,
we demonstrate that Prx V plays a key role in the
microglial activation process through modulation of the balance
between ROS/NO generation and the corresponding
JNK cascade activation.
PMID: 20345759
Keywords: C-jun N-terminal kinase; Lipopolysaccharide;
Microglia; Mutation; NADPH oxidase; Nitric oxide;
Peroxiredoxin V; Reactive oxygen species; Signal
transduction
2010 KRIBB Article Abstracts | 11 |