From FRET Imaging to Practical Methodology for Kinase Activity Sensing in Living Cells - Progress in Molecular Biology and Translational Science - page 146

JNK c-Jun N-terminal kinase
KAR kinase activity reporter
LSM least squares method
MAPK mitogen-activated protein kinase
MLE maximum likelihood estimation
MPK mitogen-activated protein kinase phosphatase
MRE molecular recognition element
PAABD phosphoamino acid-binding domain
PKA protein kinase A
PKC protein kinase C
PKG protein kinase G
PMT photomultiplier tube
PTB phosphotyrosine binding
R ratio
Rac subfamily of the Rho family of GTPases
RFP red fluorescent protein
RNA ribonucleic acid
siRNA small interfering RNA
SR sequential ratio
TCSPC time-correlated single-photon counting
TD time domain
U2OS osteosarcoma cell line
YFP yellow fluorescent protein
YPet optimized YFP for FRET

Foreword
Biological processes are intrinsically dynamic. Although traditional methods provide
valuable insights for the understanding of many biological phenomena, the possibility
of measuring, quantifying, and localizing proteins within a cell, a tissue, and even an
embryo has revolutionized our train of thoughts and has encouraged scientists to
develop molecular tools for the assessment of protein or protein complex dynamics
within their physiological context. These ongoing efforts rest on the emergence of bio-
photonic techniques and the continuous improvement of fluorescent probes, allowing
precise and reliable measurements of dynamic cellular functions. The march of the
“ in vivo biochemistry ” has begun, already yielding breathtaking results.

1. INTRODUCTION

How cells sense external and internal signals and how these signals are
processed to drive specific responses in a multiscale context are major ques-
tions in biology.
Proten phosphorylation plays a significant role in a wide range of cellular
processes such as cell proliferation, differentiation, and cellular death. In eu-
karyotes, phosphorylation occurs on serine,

threonine, tyrosine, and