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A FRET-based approach for quantitative evaluation of forskolin-induced pendrin trafficking at the plasma membrane in bronchial NCI H292 cells.
[pendred syndrome]
Human
pendrin
(
SLC
26
A
4
,
PDS
)
is
an
integral
membrane
protein
acting
as
an
electroneutral
anion
exchanger
.
Loss
of
function
mutations
in
pendrin
protein
cause
Pendred
syndrome
,
a
disorder
characterized
by
sensorineural
deafness
and
a
partial
iodide
organification
defect
that
may
lead
to
thyroid
goiter
.
Additionally
,
pendrin
up-regulation
could
play
a
role
in
the
pathogenesis
of
several
diseases
including
bronchial
asthma
and
chronic
obstructive
pulmonary
disease
(
COPD
)
.
Therefore
,
monitoring
the
plasma
membrane
abundance
and
trafficking
of
pendrin
in
the
context
of
a
living
cell
is
crucially
important
.
Trafficking
of
pendrin
to
the
plasma
membrane
was
monitored
by
fluorescence
resonance
energy
transfer
(
FRET
)
,
a
physical
phenomenon
occurring
between
two
fluorophores
(
the
FRET
donor
and
acceptor
)
located
in
close
spatial
proximity
.
Because
the
efficiency
of
the
energy
transfer
is
inversely
proportional
to
the
sixth
power
of
the
distance
between
donor
and
acceptor
,
FRET
is
extremely
sensitive
to
small
changes
in
distance
between
the
donor
and
acceptor
and
is
therefore
a
powerful
tool
to
determine
protein-protein
interactions
.
FRET
studies
revealed
that
forskolin-induced
cAMP
production
is
associated
with
a
significant
increase
of
pendrin
expression
at
plasma
membrane
,
which
is
paralleled
by
a
decrease
in
intracellular
pH
.
Pendrin
transposition
to
the
membrane
is
accompanied
with
a
partial
depolymerization
of
actin
cytoskeleton
via
Rho-
GTPase
inhibition
.
Trafficking
to
the
plasma
membrane
is
critical
in
the
regulation
of
pendrin
activity
.
Therefore
,
reliable
tools
for
monitoring
and
quantifying
this
phenomenon
are
highly
desirable
.
Diseases
Validation
Diseases presenting
"which is paralleled by a decrease in intracellular ph"
symptom
pendred syndrome
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