Ser262 determines the chloride-dependent colour tuning of a new halorhodopsin from Haloquadratum walsbyi

Hsu Yuan Fu, Yung Ning Chang, Ming Jin Jheng, Chii Shen Yang

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Light is an important environmental signal for all organisms on earth because it is essential for physiological signalling and the regulation of most biological systems. Halophiles found in salt-saturated ponds encode various archaeal rhodopsins and thereby harvest various wavelengths of light either for ion transportation or as sensory mediators. HR (halorhodopsin), one of the microbial rhodopsins, senses yellow light and transports chloride or other halides into the cytoplasm to maintain the osmotic balance during cell growth, and it exists almost ubiquitously in all known halobacteria. To date, only two HRs, isolated from HsHR (Halobacterium salinarum HR) and NpHR (Natronomonas pharaonis HR), have been characterized. In the present study, two new HRs, HmHR (Haloarcula marismortui HR) and HwHR (Haloquadratum walsbyi HR), were functionally overexpressed in Escherichia coli, and the maximum absorbance (λmax) of the purified proteins, the light-driven chloride uptake and the chloride-binding affinity were measured. The results showed them to have similar properties to two HRs reported previously. However, the λmax of HwHR is extremely consistent in a wide range of salt/chloride concentrations, which had not been observed previously. A structural-based sequence alignment identified a single serine residue at 262 in HwHR, which is typically a conserved alanine in all other known HRs. A Ser 262 to alanine replacement in HwHR eliminated the chloride-independent colour tuning, whereas an Ala262 to serine mutagenesis in HsHR transformed it to have chloride-independent colour tuning similar to that of HwHR. Thus Ser262 is a key residue for the mechanism of chloride-dependent colour tuning in HwHR.

Original languageEnglish (US)
Pages (from-to)501-509
Number of pages9
JournalBioscience Reports
Volume32
Issue number5
DOIs
StatePublished - Oct 2012

Keywords

  • Chloride affinity
  • Halorhodopsin
  • Light-driven chloride pump
  • Photocycle
  • Site-directed evolution
  • Spectral tuning

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