Association between a M-Iacking mutant D75N of pharaonis phoborhodopsin and its transducer is stronger than the complex of the wild-type pigment: Implication of the signal transduction

  • Sudo, Yuki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) ;
  • Iwamoto, Masayuki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) ;
  • Shimono, Kazumi (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) ;
  • Kamo, Naoki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University)
  • Published : 2002.08.01

Abstract

In halobacterial membrane, pharaonis phoborhodopsin (or pharaonis sensory rhdopsin II, psRII) forms a complex with its transducer pHtrII. Flash-photolyis of D75N mutant did not yield M-intermediate but an O-like intermediate is observed. We examined the interaction between D75N of ppR and t-Htr (truncated pHtrII). These formed a complex in the presence of n-dodecyl-$\beta$-D-maltoside, and the association accelerated the decay of the 0 of D75N from 15 to 56 s$\^$-1/. From the decay time constants under varying ratios of D75N and t-Htr, n, the molar ratio of D75N/t-Htr in the complex, and K$\_$D/, the dissociation constant, were estimated. The value of n was unity and K$\_$D/ was estimated to 146 nM. This K$\_$D/ value can be considered as the association between the photo-intermediate and t-Htr, which is deduced by the method of estimation. Previously we (Photochem. Photobiol. 74, 489-494 (2001)) reported K$\_$D/ of 15 $\mu$M for the interaction between the wild-type and t-Htr by means of the change of M-decay rates. Therefore, this value should be the K$\_$D/ value for the interaction between M of the wild-type and t-Htr.