From FRET Imaging to Practical Methodology for Kinase Activity Sensing in Living Cells - Progress in Molecular Biology and Translational Science - page 149

Evolution in genetically encode FRET-based biosensor

cGMP sensor

First sensor for the kinase
PKA (ART)

Metal release from
metallothionein sensor

Nobel price in chemixstry for
the discovery of the GFP

First cAMP sensor based on
GFP-tagged PKA subunits

Optimization of
PKA sensor: AKAR3

Optimization of
PKA sensor: AKAR4

First protease sensor

1996 1998 2000 2002 2004 2006 2008 2010 2012

1997 1999 2001 2003 2005 2007 2009 2011

First calcium sensor

PKA sensor:
AKAR1

Optimization of
PKA sensor: AKAR2

Creation of an optimized
generic KAR backbone Cameleon targeted to

secretory vesicle membrane

Caspase-1 and -3 sensors

Figure 5.2 Evolution in genetically encoded FRET-based biosensors.

A

Donor fluorophore Substrate PAABD Acceptor fluorophore

B

FRET

Kinase + ATP

Donor
fluorophore

phosphatase

Substrate

Acceptor
fluorophore

P

PAABD

Figure 5.3 General scheme of the functional domains of FRET-based kinase activity
reporter (KAR). The schematic representation shows the biosensor conformational
changes upon phosphorylation by the specific kinase leading to an increase in FRET be-
tween donor and acceptor fluorophores.