Role of charged residues of pharaonis phoborhodopsin (sensory rhodopsin II) in its interaction with the transducer protein

Yuki Sudo, Masayuki Iwamoto, Kazumi Shimono, Naoki Kamo

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

pharaonis phoborhodopsin (ppR; also called pharaonis sensory rhodopsin II, NpSRII) is a receptor for negative phototaxis in Natronomonas (Natronobacterium) pharaonis. In membranes, it forms a 2:2 complex with its transducer protein, pHtrII, which transmits light signals into the cytoplasmic space through protein-protein interactions. We previously found that a specific deprotonated carboxyl of ppR or pHtrII strengthens their binding [Sudo, Y., et al. (2002) Biophys. J. 83, 427-432]. In this study we aim to identify this carboxyl group. Since the D75N mutant has only one photointermediate (ppRo-like) whose existence spans the millisecond time range, the analysis of its decay rate is simple. We prepared various D75N mutants such as D75N/D214N, D75N/K157Q/R162Q/R164Q (D75N/3Gln), D75N/D193N, and D75N/D193E, among which only D75N/D193N did not show pH dependence with regard to the ppRo-like decay rate and KD value for binding, implying that the carboxyl group in question is from Asp-193. The pKa of this group decreased to below 2 when a complex was formed. Therefore, we conclude that Asp-193ppR is connected to the distant transducer-ppR binding surface via hydrogen bonds, thereby modulating its pKa. In addition, we discuss the importance of Arg-162 ppR with respect to the binding activity.

Original languageEnglish
Pages (from-to)13748-13754
Number of pages7
JournalBiochemistry
Volume43
Issue number43
DOIs
Publication statusPublished - Nov 2 2004
Externally publishedYes

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Sensory Rhodopsins
Transducers
Natronobacterium
Proteins
Hydrogen
Hydrogen bonds
Membranes
Light

ASJC Scopus subject areas

  • Biochemistry

Cite this

Role of charged residues of pharaonis phoborhodopsin (sensory rhodopsin II) in its interaction with the transducer protein. / Sudo, Yuki; Iwamoto, Masayuki; Shimono, Kazumi; Kamo, Naoki.

In: Biochemistry, Vol. 43, No. 43, 02.11.2004, p. 13748-13754.

Research output: Contribution to journalArticle

Sudo, Yuki ; Iwamoto, Masayuki ; Shimono, Kazumi ; Kamo, Naoki. / Role of charged residues of pharaonis phoborhodopsin (sensory rhodopsin II) in its interaction with the transducer protein. In: Biochemistry. 2004 ; Vol. 43, No. 43. pp. 13748-13754.
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