Effect of chloride binding on the thermal trimer-monomer conversion of halorhodopsin in the solubilized system

Biochemistry
Takanori SasakiMakoto Demura

Abstract

Halorhodopsin from Natronomonas pharaonis (NpHR) acts an inward-directed, light-driven chloride pump and forms a homotrimer. To evaluate effect of trimeric assembly, that is, intermolecular interaction, on the control or modulation of light-driven chloride pumping activity of individual HRs, it is important to understand the thermal and chloride sensitivity of trimer dissociation and the structural stability of HR. In this study, the thermal dissociation of NpHR trimer to monomer in a dodecyl beta-d-maltoside-solubilized system was investigated, using size-exclusion chromatography and visible absorption. In the absence of Cl(-), NpHR retained the trimer assembly at 25 degrees C but dissociated to the monomer with an increase in temperature to >40 degrees C. On the other hand, in the presence of Cl(-), the trimer assembly was maintained at 40 degrees C. The dissociation of the trimer to the monomer after incubation at 40 degrees C, which was determined via size-exclusion chromatography, depended on the Cl(-) concentration and showed a sigmoidal isotherm. From this isotherm, the apparent dissociation constant for Cl(-) was estimated to be 22 mM with a Hill coefficient of 2.2. A similar isotherm was obtained when SO(4)(2-) was use...Continue Reading

References

Jan 1, 1980·Archives of Biochemistry and Biophysics·A Matsuno-Yagi, Y Mukohata
Jul 2, 2003·Biophysical Chemistry·Maki SatoKatsutoshi Nitta
Aug 29, 2003·Journal of Biochemistry·Maki SatoKatsutoshi Nitta
Jun 7, 2005·Photochemistry and Photobiology·Takanori SasakiShigeki Mitaku
Jul 19, 2005·Journal of Molecular Biology·Hideo OkumuraTsutomu Kouyama
Nov 23, 2006·Photochemistry and Photobiology·Walter GmelinDieter Oesterhelt
Apr 6, 2007·Nature·Feng ZhangKarl Deisseroth
Mar 10, 2009·Photochemistry and Photobiology·Megumi KuboMakoto Demura
May 6, 2009·Biophysical Journal·Keiichi InoueMasahide Terazima

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Citations

Apr 25, 2015·Biochemistry·Sansa DuttaMordechai Sheves
Jun 28, 2012·Biophysical Journal·Takashi TsukamotoMakoto Demura
Oct 4, 2014·The Journal of Physical Chemistry. B·Takashi TsukamotoYuki Sudo
May 13, 2020·Biophysical Journal·Ayumi YamamotoKoichiro Ishimori
Jun 13, 2018·The Journal of Physical Chemistry. B·Tomomi ShionoyaYasuhisa Mizutani

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