PMID- 29494125
OWN - NLM
STAT- MEDLINE
DCOM- 20190417
LR  - 20190417
IS  - 1554-8937 (Electronic)
IS  - 1554-8929 (Linking)
VI  - 13
IP  - 7
DP  - 2018 Jul 20
TI  - Bioluminescent Low-Affinity Ca(2+) Indicator for ER with Multicolor Calcium 
      Imaging in Single Living Cells.
PG  - 1862-1871
LID - 10.1021/acschembio.7b01014 [doi]
AB  - The sarco/endoplasmic reticulum (SR/ER) is the foremost intercellular Ca(2+) 
      store (at submillimolar concentrations), playing a crucial role in controlling 
      intracellular Ca(2+) levels. For the investigation of SR/ER Ca(2+) dynamics in 
      cells, fluorescent protein-based genetically encoded calcium indicators (GECIs) 
      with low Ca(2+) affinity have been used. Recently, bioluminescent protein-based 
      GECIs with high brightness have been reported to counter the constraints of 
      fluorescence imaging, such as phototoxicity. However, their Ca(2+) affinity is 
      high and limited for imaging in the cytosol, nucleus, or mitochondria. In this 
      study, we developed a novel cyan color, low-affinity ( K(d) = 110 muM) 
      intensiometric bioluminescent GECI, which enables monitoring of the Ca(2+) 
      dynamics in the ER of HeLa cells and the SR of C2C12-derived myotubes. To 
      facilitate the broad concentration range of Ca(2+) in cellular organelles, we 
      additionally developed an intermediate affinity ( K(d) = 18 muM), orange color, 
      and bioluminescent GECI, which enables monitoring of Ca(2+) dynamics in the 
      mitochondria of HeLa cells. With these indicators, in conjunction with an 
      existing high-affinity, green, bioluminescent GECI, we succeeded in multicolor 
      bioluminescent Ca(2+) imaging in three distinct organelles (nuclei, mitochondria, 
      and ER) simultaneously. The multicolor, live, bioluminescent Ca(2+) imaging 
      demonstrated here can be used to stably reveal the ER Ca(2+) homeostasis and 
      cooperative Ca(2+) regulation among organelles. This will lead to the further 
      understanding of Ca(2+)-related physiological functions and pathophysiological 
      mechanisms.
FAU - Hossain, Md Nadim
AU  - Hossain MN
AUID- ORCID: 0000-0002-8086-8939
AD  - Department of Biotechnology, Graduate School of Engineering , Osaka University , 
      2-1 Yamadaoka , Suita 565-0871 , Japan.
FAU - Suzuki, Kazushi
AU  - Suzuki K
AD  - Department of Biotechnology, Graduate School of Engineering , Osaka University , 
      2-1 Yamadaoka , Suita 565-0871 , Japan.
FAU - Iwano, Megumi
AU  - Iwano M
AD  - Department of Biomolecular Science and Engineering, The Institute of Scientific 
      and Industrial Research , Osaka University , 8-1 Mihogaoka , Ibaraki 567-0047 , 
      Japan.
FAU - Matsuda, Tomoki
AU  - Matsuda T
AD  - Department of Biotechnology, Graduate School of Engineering , Osaka University , 
      2-1 Yamadaoka , Suita 565-0871 , Japan.
AD  - Department of Biomolecular Science and Engineering, The Institute of Scientific 
      and Industrial Research , Osaka University , 8-1 Mihogaoka , Ibaraki 567-0047 , 
      Japan.
FAU - Nagai, Takeharu
AU  - Nagai T
AD  - Department of Biotechnology, Graduate School of Engineering , Osaka University , 
      2-1 Yamadaoka , Suita 565-0871 , Japan.
AD  - Department of Biomolecular Science and Engineering, The Institute of Scientific 
      and Industrial Research , Osaka University , 8-1 Mihogaoka , Ibaraki 567-0047 , 
      Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180309
PL  - United States
TA  - ACS Chem Biol
JT  - ACS chemical biology
JID - 101282906
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Luminescent Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/*metabolism
MH  - Cell Nucleus/metabolism
MH  - Color
MH  - Endoplasmic Reticulum/*metabolism
MH  - Fluorescence
MH  - Fluorescent Dyes/*chemistry
MH  - HeLa Cells
MH  - Humans
MH  - Luminescent Proteins/*chemistry/genetics
MH  - Mice
MH  - Microscopy, Fluorescence/methods
MH  - Mitochondria/metabolism
MH  - Muscle Fibers, Skeletal/metabolism
MH  - Mutation
MH  - Sarcoplasmic Reticulum/metabolism
EDAT- 2018/03/02 06:00
MHDA- 2019/04/18 06:00
CRDT- 2018/03/02 06:00
PHST- 2018/03/02 06:00 [pubmed]
PHST- 2019/04/18 06:00 [medline]
PHST- 2018/03/02 06:00 [entrez]
AID - 10.1021/acschembio.7b01014 [doi]
PST - ppublish
SO  - ACS Chem Biol. 2018 Jul 20;13(7):1862-1871. doi: 10.1021/acschembio.7b01014. Epub 
      2018 Mar 9.