• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.20 no.1
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• pp.80-85
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• 2007

We report the electro-optical properties of the sensory rhodopsin II using a near-field scanning microwave microscope(NSMM). Rhodopsin was known as a photoreceptor pigment with a retinal as a chromophore via a protonated Schiff base and consists of seven ${\alpha}-helical$ transmembrane segments. The sensory rhodopsin II, expressing E. coli UT5600 with endogenous retinal biosynthesis system and purified with $Ni^{-2}-NTA$ affinity chromatography in the presence of 0.02 % DM (Dodecyl Maltoside) from Natronomonas pharaonis. We measured the absorption spectra and the transients difference of sensory rhodopsin II from Natronomonas pharaonis using a UV/VIS spectrophotometer with Nd-Yag Laser (532 nm). The absorption spectra of NpSR II showed a typical rhodopsin spectrum with a left shoulder region and the photointermediates spectra of NpSR II-ground state (${\lambda}max=498\;nm$), NpSR II-M state (${\lambda}max=390\;nm$), and NpSR II-O state (${\lambda}max=550\;nm$) during the photocycle. The observed photocycle reaction was confirmed by measuring the microwave reflection coefficient $S_{11}$ at an operating frequency of f=3.93-3.95 GHz and compared with the results of a photocycle of NpSR II.

• Journal of Photoscience
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• v.9 no.3
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• pp.45-48
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• 2002

Microbial rhodopsins, photoactive 7-transmembrane helix proteins that use retinal as their chromophore, were observed initially in the Archaea and appeared to be restricted to extreme halophilic environments. Our understanding of the abundance and diversity of this family has been radically transformed by findings over the past three years. Genome sequencing of cultivated microbes as well as environmental genomics have unexpectedly revealed archaeal rhodopsin homologs in the other two domains of life as well, namely Bacteria and Eucarya. Organisms containing these homologs inhabit such diverse environments as salt flats, soil, freshwater, and surface and deep ocean waters, and they comprise a broad phylogenetic range of microbial life, including haloarchaea, proteobacteria, cyanobacteria, fungi, and algae. Analysis of the new microbial rhodopsins and their expression and structural and functional characterization reveal that they fulfill both ion transport and sensory functions in various organisms, and use a variety of signaling mechanisms. We have obtained the first crystallographic structure for a photosensory member of this family, the phototaxis receptor sensory rhodopsin II (SRII, also known as phoborhodopsin) that mediates blue-light avoidance by the haloarchaeon Natronobacterium pharaonis. The structure obtained from x-ray diffraction of 3D crystals prepared in a cubic lipid phase reveals key features responsible for its spectral tuning and its sensory function. The mechanism of SRII signaling fits a unified model for transport and signaling in this widespread family of phototransducers.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.22-29
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• 2011

Rhodopsin is a membrane protein with seven transmembrane region which contains a retinal as its chromophore. Although there have been recently reports on various photo-biochemical features of rhodopsins by a wide range of purifying and measurement methods, there was no actual comparison related to the difference of biochemical characteristics according to their physical environment of rhodopsins. First, proteorhodopsin (PR) was found in marine proteobacteria whose function is known for pumping proton using light energy. Second one is Anabaena sensory rhodopsin (Nostoc sp.) PCC7120 (ASR) which belongs to eubacteria acts as sensory regulator since it is co-expressed with transducer 14 kDa in an operon. In this study, we applied two types of rhodopsins (PR and ASR) to various environmental conditions such as in Escherichia coli membranes, membrane in acrylamide gel, in DDM (n-dodecyl-${\beta}$-D-maltopyranoside), OG (octyl-${\beta}$-D-glucopyranoside), and reconstituted with DOPC (1,2-didecanoyl-sn-glycero-3-phosphocholine). According to the light-induced difference spectroscopy, rhodopsins in 0.02% DDM clearly showed photointermediates like M, and O states which respond to the different wavelengths, respectively and showed the best signal/noise ratio. The laser-induced difference spectra showed the fast formation and decay rate of photointermediates in the DDM solubilized samples than gel encapsulated rhodopsin. Each of rhodopsins seemed to be adapted to its surrounding environment.

• Journal of Photoscience
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• v.9 no.2
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• pp.5-8
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• 2002

Photoperiodism is an important adaptive phenomenon in various physiological parameters including reproduction to cope with seasonal changes. Involvement of extraretinal photoreceptors in the photoperiodism in non-mammalian vertebrates has been well established. In addition, circadian clock system is known to be involved in the photoperiodic time measurement. The pathway consists of light-input system, time measurement system (circadian clock), gonadotropin releasing hormone (GnRH) production in the hypothalamus, luteinizing hormone (LH) and follicle stimulating hormone (FSH) production in the pituitary, and final gonadal development. Recently, several laboratories reported photopigments newly cloned in the pineal, eyes and deep brain in addition to already known visual pigments in the retina. These are pinopsin, parapinopsin, VA-opsin, melanopsin, etc. All these photopigments belong to the opsin family having retinal as the chromophore. However, the function of these photopigments remains unknown. I reviewed the studies on the location of the photopigments by immunocytochemistry. I also discussed the results on the action spectra for induction of gonadal development in relation with the location of the photoreceptors. Various physiologically active substances distribute in the vertebrate brain. Such substances are GnRH, GnIH, neuropeptide Y, vasoactive intestinal peptide, c-Fos, galanin, neurosteroids, etc. I summarized the immunhistochemical studies on the distribution and the photoperiodic changes of these substances and discussed the route from the deep brain photoreceptor to GnRH cells.

• Journal of Photoscience
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• v.9 no.2
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• pp.41-43
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• 2002

In photoreceptor cells, light activates visual pigments consisting of a chromophore (retinal) and a protein moiety (opsin). Activated visual pigments trigger an enzymatic cascade, called phototransduction cascade, in which more than ten phototransduction proteins are participating. Two types of vertebrate photoreceptor cells, rods and cones, play roles in twilight and daylight vision, respectively. Cones are further classified into several subtypes based on their morphology and spectral sensitivity. Though the diversities of vertebrate photoreceptor cells are crucial for color discrimination and detection of light over a wider range of intensities, the molecular mechanism to characterize the photoreceptor types remains unclear. We investigated the amino acid sequences of about 50 vertebrate opsins, and found that these sequences can be classified into five fundamental subfamilies. Clear relationships were found between these subfamilies and their characteristic spectral sensitivities. In addition to opsins, we studied other phototransduction proteins. The amino acid sequences of phototransduction proteins can be classified into a few subfamilies. Even though their spectral sensitivity is considerably different, cones fundamentally share the phototransduction protein isoforms which are different from those found in rods. It is suggested that the difference in phototransduction proteins between rods and cones is responsible for their sensitivity to light. Isoforms and their selective expression may characterize individual photoreceptor cells, thus providing us with physiological functions such as color vision and daylight/twilight visions.