Open Access
Scientific Reports, volume 8, issue 1, publication number 15542
Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids
Solovyev Ilya D
1, 2
,
Gavshina Alexandra V
1
,
Katti Aditya S
3
,
Chizhik Alexey I
3
,
Vinokurov Leonid M.
4
,
Lapshin Grigory D
1
,
Ivashina Tatiana V
5
,
Khrenova Maria G
1, 2
,
Kireev I.
6
,
Gregor Ingo
3
,
Enderlein Jörg
3
,
Savitsky Alexander
1, 2
1
3
University of Göttingen, Third Institute of Physics – Biophysics, Göttingen, Germany
|
Publication type: Journal Article
Publication date: 2018-10-19
PubMed ID:
30341334
Multidisciplinary
Abstract
Photoconvertible fluorescent proteins (PCFPs) are widely used as markers for the visualization of intracellular processes and for sub-diffraction single-molecule localization microscopy. Although wild type of a new photoconvertible fluorescent protein SAASoti tends to aggregate, we succeeded, via rational mutagenesis, to obtain variants that formed either tetramers or monomers. We compare two approaches: one is based on the structural similarity between SAASoti and Kaede, which helped us to identify a single point mutation (V127T) at the protein’s hydrophobic interface that leads to monomerization. The other is based on a chemical modification of amino groups of SAASoti with succinic anhydride, which converts the protein aggregates into monomers. Mass-spectrometric analysis helped us to identify that the modification of a single ε-amino group of lysine K145 in the strongly charged interface AB was sufficient to convert the protein into its tetrameric form. Furthermore, site-directed mutagenesis was used to generate mutants that proved to be either monomeric or tetrameric, both capable of rapid green-to-red photoconversion. This allows SAASoti to be used as a photoconvertible fluorescent marker for in vivo cell studies.
Citations by journals
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1
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3
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Scientific Reports
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Scientific Reports
3 publications, 37.5%
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International Journal of Molecular Sciences
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International Journal of Molecular Sciences
2 publications, 25%
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Materials
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Materials
1 publication, 12.5%
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RSC Chemical Biology
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RSC Chemical Biology
1 publication, 12.5%
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Applied Biochemistry and Microbiology
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Applied Biochemistry and Microbiology
1 publication, 12.5%
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1
2
3
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Citations by publishers
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1
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3
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Springer Nature
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Springer Nature
3 publications, 37.5%
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Multidisciplinary Digital Publishing Institute (MDPI)
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Multidisciplinary Digital Publishing Institute (MDPI)
3 publications, 37.5%
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Royal Society of Chemistry (RSC)
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Royal Society of Chemistry (RSC)
1 publication, 12.5%
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Pleiades Publishing
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Pleiades Publishing
1 publication, 12.5%
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1
2
3
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- We do not take into account publications that without a DOI.
- Statistics recalculated only for publications connected to researchers, organizations and labs registered on the platform.
- Statistics recalculated weekly.
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Solovyev I. D. et al. Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids // Scientific Reports. 2018. Vol. 8. No. 1. 15542
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Solovyev I. D., Gavshina A. V., Katti A. S., Chizhik A. I., Vinokurov L. M., Lapshin G. D., Ivashina T. V., Khrenova M. G., Kireev I., Gregor I., Enderlein J., Savitsky A. Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids // Scientific Reports. 2018. Vol. 8. No. 1. 15542
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TY - JOUR
DO - 10.1038/s41598-018-33250-z
UR - https://doi.org/10.1038%2Fs41598-018-33250-z
TI - Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids
T2 - Scientific Reports
AU - Solovyev, Ilya D
AU - Gavshina, Alexandra V
AU - Katti, Aditya S
AU - Chizhik, Alexey I
AU - Vinokurov, Leonid M.
AU - Lapshin, Grigory D
AU - Ivashina, Tatiana V
AU - Khrenova, Maria G
AU - Kireev, I.
AU - Gregor, Ingo
AU - Enderlein, Jörg
AU - Savitsky, Alexander
PY - 2018
DA - 2018/10/19 00:00:00
PB - Springer Nature
IS - 1
VL - 8
PMID - 30341334
SN - 2045-2322
ER -
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@article{2018_Solovyev,
author = {Ilya D Solovyev and Alexandra V Gavshina and Aditya S Katti and Alexey I Chizhik and Leonid M. Vinokurov and Grigory D Lapshin and Tatiana V Ivashina and Maria G Khrenova and I. Kireev and Ingo Gregor and Jörg Enderlein and Alexander Savitsky},
title = {Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids},
journal = {Scientific Reports},
year = {2018},
volume = {8},
publisher = {Springer Nature},
month = {oct},
url = {https://doi.org/10.1038%2Fs41598-018-33250-z},
number = {1},
doi = {10.1038/s41598-018-33250-z}
}