Importance of specific hydrogen bonds of archaeal rhodopsins for the binding to the transducer protein

Yuki Sudo, Masaki Yamabi, Shinnosuke Kato, Chisa Hasegawa, Masayuki Iwamoto, Kazumi Shimono, Naoki Kamo

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Four rhodopsins, bacteriorhodopsin (bR), halorhodopsin (hR), sensory rhodopsin (sR) and phoborhodopsin (pR) exist in archaeal membranes. bR and hR work as a light-driven ion pump. sR and pR work as a photo-sensor of phototaxis, and form signaling complexes in membranes with their respective cognate transducer proteins HtrI (with sR) and HtrII (with pR), through which light signals are transmitted to the cytoplasm. What is the determining factor(s) of the specific binding to form the complex? Binding of the wild-type or mutated rhodopsins with HtrII was measured by isothermal titration calorimetric analysis (ITC). bR and hR could not bind with HtrII. On the other hand, sR could bind to HtrII, although the dissociation constant (K D) was about 100 times larger than that of pR. An X-ray crystallographic structure of the pR/HtrII complex revealed formation of two specific hydrogen bonds whose pairs are Tyr199 pR/Asn74 HtrII and Thr189 pR/Glu43 HtrII/Ser62 HtrII. To investigate the importance of these hydrogen bonds, the K D value for the binding of various mutants of bR, hR, sR and pR with HtrII was estimated by ITC. The K D value of T189V pR/Y199F pR, double mutant/HtrII complex, was about 100-fold larger than that of the wild-type pR, whose K D value was 0.16 μM. On the other hand, bR and hR double mutants, P200T bR/V210Y bR and P240T hR/F250Y hR, were able to bind with HtrII. The K D value of these complexes was estimated to be 60.1(±10.7) μM for bR and to be 29.1(±6.1) μM for hR, while the wild-type bR and hR did not bind with HtrII. We concluded that these two specific hydrogen bonds play important roles in the binding between the rhodopsins and transducer protein.

Original languageEnglish
Pages (from-to)1274-1282
Number of pages9
JournalJournal of Molecular Biology
Volume357
Issue number4
DOIs
Publication statusPublished - Apr 7 2006
Externally publishedYes

Fingerprint

Microbial Rhodopsins
Halorhodopsins
Bacteriorhodopsins
Transducers
Sensory Rhodopsins
Hydrogen
Proteins
Rhodopsin
Pair Bond
Ion Pumps
Light
Membranes
Cytoplasm

Keywords

  • Ion pump
  • Phoborhodopsin
  • Photo-sensor
  • Protein-protein interaction
  • Sensory rhodopsin

ASJC Scopus subject areas

  • Virology

Cite this

Importance of specific hydrogen bonds of archaeal rhodopsins for the binding to the transducer protein. / Sudo, Yuki; Yamabi, Masaki; Kato, Shinnosuke; Hasegawa, Chisa; Iwamoto, Masayuki; Shimono, Kazumi; Kamo, Naoki.

In: Journal of Molecular Biology, Vol. 357, No. 4, 07.04.2006, p. 1274-1282.

Research output: Contribution to journalArticle

Sudo, Yuki ; Yamabi, Masaki ; Kato, Shinnosuke ; Hasegawa, Chisa ; Iwamoto, Masayuki ; Shimono, Kazumi ; Kamo, Naoki. / Importance of specific hydrogen bonds of archaeal rhodopsins for the binding to the transducer protein. In: Journal of Molecular Biology. 2006 ; Vol. 357, No. 4. pp. 1274-1282.
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AU - Shimono, Kazumi

AU - Kamo, Naoki

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