TY - JOUR
T1 - Concentration Quantification of the Low-Complexity Domain of Fused in Sarcoma inside a Single Droplet and Effects of Solution Parameters
AU - Yokosawa, Kohei
AU - Kajimoto, Shinji
AU - Shibata, Daiki
AU - Kuroi, Kunisato
AU - Konno, Tomohiro
AU - Nakabayashi, Takakazu
N1 - Funding Information:
This work was supported by JSPS KAKENHI Grant Numbers JP20K21468 (T.N.), JP20H04689 (S.K.), and JP19H02666 (S.K.) from the Ministry of Education, Culture, Sports, Science and Technology in Japan; JST PRESTO Grant Number JPMJPR20E5 (S.K.) in Japan; and a grant of Joint Research by the National Institutes of Natural Sciences (NINS) (NINS program 01112103). We thank Prof. Naoto Ueno for the opportunity of NINS Joint Research.
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/6/23
Y1 - 2022/6/23
N2 - Liquid-liquid phase separation (LLPS) is an important phenomenon in biology, and it is desirable to develop quantitative methods to analyze protein droplets generated by LLPS. This study quantified the change in protein concentration in a droplet in label-free and single-droplet conditions using Raman imaging and the Raman band of water as an intensity standard. Small changes in the protein concentration with variations in pH and salt concentration were observed, and it was shown that the concentration in the droplet decreases as the conditions become less favorable for droplet formation. The effect of exposure to 1,6-hexanediol was also examined, and this additive was found to decrease the protein concentration in the droplet. A model can be proposed in which the addition of 1,6-hexanediol reduces the protein concentration in the droplet, and the droplet disappears when the concentration falls below a certain threshold value.
AB - Liquid-liquid phase separation (LLPS) is an important phenomenon in biology, and it is desirable to develop quantitative methods to analyze protein droplets generated by LLPS. This study quantified the change in protein concentration in a droplet in label-free and single-droplet conditions using Raman imaging and the Raman band of water as an intensity standard. Small changes in the protein concentration with variations in pH and salt concentration were observed, and it was shown that the concentration in the droplet decreases as the conditions become less favorable for droplet formation. The effect of exposure to 1,6-hexanediol was also examined, and this additive was found to decrease the protein concentration in the droplet. A model can be proposed in which the addition of 1,6-hexanediol reduces the protein concentration in the droplet, and the droplet disappears when the concentration falls below a certain threshold value.
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U2 - 10.1021/acs.jpclett.2c00962
DO - 10.1021/acs.jpclett.2c00962
M3 - Article
C2 - 35709358
AN - SCOPUS:85133100765
SN - 1948-7185
VL - 13
SP - 5692
EP - 5697
JO - Journal of Physical Chemistry Letters
JF - Journal of Physical Chemistry Letters
IS - 24
ER -