PMID- 31427680
OWN - NLM
STAT- MEDLINE
DCOM- 20201021
LR  - 20210110
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Aug 19
TI  - ShadowR: a novel chromoprotein with reduced non-specific binding and improved 
      expression in living cells.
PG  - 12072
LID - 10.1038/s41598-019-48604-4 [doi]
LID - 12072
AB  - Here we developed an orange light-absorbing chromoprotein named ShadowR as a 
      novel acceptor for performing fluorescence lifetime imaging microscopy-based 
      Forster resonance energy transfer (FLIM-FRET) measurement in living cells. 
      ShadowR was generated by replacing hydrophobic amino acids located at the surface 
      of the chromoprotein Ultramarine with hydrophilic amino acids in order to reduce 
      non-specific interactions with cytosolic proteins. Similar to Ultramarine, 
      ShadowR shows high absorption capacity and no fluorescence. However, it exhibits 
      reduced non-specific binding to cytosolic proteins and is highly expressed in 
      HeLa cells. Using tandem constructs and a LOVTRAP system, we showed that ShadowR 
      can be used as a FRET acceptor in combination with donor mRuby2 or mScarlet in 
      HeLa cells. Thus, ShadowR is a useful, novel FLIM-FRET acceptor.
FAU - Murakoshi, Hideji
AU  - Murakoshi H
AD  - Supportive Center for Brain Research, National Institute for Physiological 
      Sciences, Okazaki, Aichi, 444-8585, Japan. murakosh@nips.ac.jp.
AD  - Department of Physiological Sciences, The Graduate University for Advanced 
      Studies, Hayama, Kanagawa, 240-0193, Japan. murakosh@nips.ac.jp.
FAU - Horiuchi, Hiroshi
AU  - Horiuchi H
AD  - Department of Physiological Sciences, The Graduate University for Advanced 
      Studies, Hayama, Kanagawa, 240-0193, Japan.
AD  - Division of Homeostatic Development, National Institute for Physiological 
      Sciences, Okazaki, Aichi, 444-8585, Japan.
FAU - Kosugi, Takahiro
AU  - Kosugi T
AUID- ORCID: 0000-0002-6289-5319
AD  - Department of Structural Molecular Science, The Graduate University for Advanced 
      Studies, Hayama, Kanagawa, 240-0193, Japan.
AD  - Exploratory Research Center on Life and Living Systems (ExCELLS), Okazaki, Aichi, 
      444-8585, Japan.
AD  - Research Center of Integrative Molecular Systems, Institute for Molecular 
      Science, Okazaki, Aichi, 444-8585, Japan.
FAU - Onda, Maki
AU  - Onda M
AD  - Supportive Center for Brain Research, National Institute for Physiological 
      Sciences, Okazaki, Aichi, 444-8585, Japan.
FAU - Sato, Aiko
AU  - Sato A
AD  - Supportive Center for Brain Research, National Institute for Physiological 
      Sciences, Okazaki, Aichi, 444-8585, Japan.
FAU - Koga, Nobuyasu
AU  - Koga N
AD  - Department of Structural Molecular Science, The Graduate University for Advanced 
      Studies, Hayama, Kanagawa, 240-0193, Japan.
AD  - Exploratory Research Center on Life and Living Systems (ExCELLS), Okazaki, Aichi, 
      444-8585, Japan.
AD  - Research Center of Integrative Molecular Systems, Institute for Molecular 
      Science, Okazaki, Aichi, 444-8585, Japan.
FAU - Nabekura, Junichi
AU  - Nabekura J
AD  - Department of Physiological Sciences, The Graduate University for Advanced 
      Studies, Hayama, Kanagawa, 240-0193, Japan.
AD  - Division of Homeostatic Development, National Institute for Physiological 
      Sciences, Okazaki, Aichi, 444-8585, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190819
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Luminescent Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - *Biophysical Phenomena
MH  - *Fluorescence
MH  - Fluorescence Resonance Energy Transfer
MH  - Gene Expression/genetics
MH  - Green Fluorescent Proteins/chemistry/genetics
MH  - HeLa Cells
MH  - Humans
MH  - Luminescent Proteins/*chemistry/genetics
MH  - Microscopy, Fluorescence/*methods
MH  - Protein Binding/genetics
PMC - PMC6700193
COIS- The authors declare no competing interests.
EDAT- 2019/08/21 06:00
MHDA- 2020/10/22 06:00
PMCR- 2019/08/19
CRDT- 2019/08/21 06:00
PHST- 2019/02/12 00:00 [received]
PHST- 2019/08/08 00:00 [accepted]
PHST- 2019/08/21 06:00 [entrez]
PHST- 2019/08/21 06:00 [pubmed]
PHST- 2020/10/22 06:00 [medline]
PHST- 2019/08/19 00:00 [pmc-release]
AID - 10.1038/s41598-019-48604-4 [pii]
AID - 48604 [pii]
AID - 10.1038/s41598-019-48604-4 [doi]
PST - epublish
SO  - Sci Rep. 2019 Aug 19;9(1):12072. doi: 10.1038/s41598-019-48604-4.