Structural analysis of the phototactic transducer protein HtrII linker region from Natronomonas pharaonis

Kokoro Hayashi, Yuki Sudo, JunGoo Jee, Masaki Mishima, Hideyuki Hara, Naoki Kamo, Chojiro Kojima

Research output: Contribution to journalArticle

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Abstract

Halobacterial pharaonis phoborhodopsin [ppR, also called Natronomonas pharaonis sensory rhodopsin II (NpSRII)] is a phototaxis protein which transmits a light signal to the cytoplasm through its transducer protein (pHtrII). pHtrII, a two-transmembrane protein that interacts with ppR, belongs to the group of methyl-accepting chemotaxis proteins (MCPs). Several mutation studies have indicated that the linker region connecting the transmembrane and methylation regions is necessary for signal transduction. However, the three-dimensional (3D) structure of an MCP linker region has yet to be reported, and hence, details concerning the signal transduction mechanism remain unknown. Here the structure of the pHtrII linker region was investigated biochemically and biophysically. Following limited proteolysis, only one trypsin resistant fragment in the pHtrII linker region was identified. This fragment forms a homodimer with a Kd value of 115 μM. The 3D structure of this fragment was determined by solution NMR, and only one α-helix was found between two HAMP domains of the linker region. This α-helix was significantly stabilized within transmembrane protein pHtrII as revealed by CW-EPR. The presence of Af1503 HAMP domain-like structures in the linker region was supported by CD, NMR, and ELDOR data. The α-helix determined here presumably works as a mechanical joint between two HAMP domains in the linker region to transfer the photoactivated conformational change downstream.

Original languageEnglish
Pages (from-to)14380-14390
Number of pages11
JournalBiochemistry
Volume46
Issue number50
DOIs
Publication statusPublished - Dec 18 2007
Externally publishedYes

Fingerprint

Transducers
Structural analysis
Signal Transduction
Signal transduction
Sensory Rhodopsins
Proteins
Nuclear magnetic resonance
Trypsin
Proteolysis
Methylation
Cytoplasm
Joints
Light
Mutation
Paramagnetic resonance
Methyl-Accepting Chemotaxis Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural analysis of the phototactic transducer protein HtrII linker region from Natronomonas pharaonis. / Hayashi, Kokoro; Sudo, Yuki; Jee, JunGoo; Mishima, Masaki; Hara, Hideyuki; Kamo, Naoki; Kojima, Chojiro.

In: Biochemistry, Vol. 46, No. 50, 18.12.2007, p. 14380-14390.

Research output: Contribution to journalArticle

Hayashi, K, Sudo, Y, Jee, J, Mishima, M, Hara, H, Kamo, N & Kojima, C 2007, 'Structural analysis of the phototactic transducer protein HtrII linker region from Natronomonas pharaonis', Biochemistry, vol. 46, no. 50, pp. 14380-14390. https://doi.org/10.1021/bi701837n
Hayashi, Kokoro ; Sudo, Yuki ; Jee, JunGoo ; Mishima, Masaki ; Hara, Hideyuki ; Kamo, Naoki ; Kojima, Chojiro. / Structural analysis of the phototactic transducer protein HtrII linker region from Natronomonas pharaonis. In: Biochemistry. 2007 ; Vol. 46, No. 50. pp. 14380-14390.
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