Tyr-199 and charged residues of pharaonis phoborhodopsin are important for the interaction with its transducer

Yuki Sudo, Masayuki Iwamoto, Kazumi Shimono, Naoki Kamo

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

pharaonis Phoborhodopsin (ppR; also pharaonis sensory rhodopsin II, psRII) is a retinal protein in Natronobacterium pharaonis and is a receptor of negative phototaxis. It forms a complex with its transducer, pHtrII, in membranes and transmits light signals by protein-protein interaction. Tyr-199 is conserved completely in phoborhodopsins among a variety of archaea, but it is replaced by Val (for bacteriorhodopsin) and Phe (for sensory rhodopsin I). Previously, we (Sudo, Y., M. Iwamoto, K. Shimono, and N. Kamo, submitted for publication) showed that analysis of flash-photolysis data of a complex between D75N and the truncated pHtrII (t-Htr) give a good estimate of the dissociation constant KD in the dark. To investigate the importance of Tyr-199, KD of double mutants of D75N/Y199F or D75N/Y199V with t-Htr was estimated by flash-photolysis and was ∼10-fold larger than that of D75N, showing the significant contribution of Tyr-199 to binding. The KD of the D75N/t-Htr complex increased with decreasing pH, and the data fitted well with the Henderson-Hasselbach equation with a single pKa of 3.86 ± 0.02. This suggests that certain deprotonated carboxyls at the surface of the transducer (possibly Asp-102, Asp-104, and Asp-106) are needed for the binding.

Original languageEnglish
Pages (from-to)427-432
Number of pages6
JournalBiophysical Journal
Volume83
Issue number1
Publication statusPublished - 2002
Externally publishedYes

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Sensory Rhodopsins
Transducers
Photolysis
Natronobacterium
Bacteriorhodopsins
Proteins
Archaea
Publications
Light
Membranes

ASJC Scopus subject areas

  • Biophysics

Cite this

Tyr-199 and charged residues of pharaonis phoborhodopsin are important for the interaction with its transducer. / Sudo, Yuki; Iwamoto, Masayuki; Shimono, Kazumi; Kamo, Naoki.

In: Biophysical Journal, Vol. 83, No. 1, 2002, p. 427-432.

Research output: Contribution to journalArticle

Sudo, Yuki ; Iwamoto, Masayuki ; Shimono, Kazumi ; Kamo, Naoki. / Tyr-199 and charged residues of pharaonis phoborhodopsin are important for the interaction with its transducer. In: Biophysical Journal. 2002 ; Vol. 83, No. 1. pp. 427-432.
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