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

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Title: Tyr-199 and charged residues of pharaonis Phoborhodopsin are important for the interaction with its transducer.
Authors: Sudo, Yuki Browse this author
Iwamoto, Masayuki Browse this author
Shimono, Kazumi Browse this author
Kamo, Naoki Browse this author
Keywords: Sensory rhodopsin
protein-protein interaction
Mintermediate
Halobacterium salinarum
Natronobacterium pharaonis
Issue Date: Jul-2002
Publisher: The Biophysical Society
Journal Title: Biophysical Journal
Volume: 83
Issue: 1
Start Page: 427
End Page: 432
PMID: 12080131
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.
Type: article (author version)
URI: http://hdl.handle.net/2115/16002
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 加茂 直樹

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