• Journal of Photoscience
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• v.9 no.2
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• pp.102-105
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• 2002

Phoborhodopsin (pR or sensory rhodopsin II, sRII; the absorption maximum of ∼ 500 nm) is a retinoid protein and works as a photoreceptor of the negative phototaxis of Halobacterium salinarum. pharaonis phoborhodopsin (ppR or pharaonis sensory rhodopsin II, psRII) is a corresponding protein of Natronobacterium pharaonis. These sensory proteins form a complex with a cognate transducer protein in the membrane, and this complex transmits the light-signal to the cytoplasm to evoke avoidance reaction from blue-green light. Recently, the functional expression in Escherichia coli membrane of ppR was achieved, which can afford a large amount of the protein and enables mutant studies to clarify the role of various amino acid residues. A truncated transducer which can bind to ppR is also expressed in Escherichia. coli membrane. In this article, we will review properties of ppR mainly using observations of our laboratory; which contains photochemistry (photocycle), light-driven proton uptake, release and transport, F -helix titling during photocycle and association of the transducer.

• Journal of Photoscience
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• v.4 no.3
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• pp.133-140
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• 1997

A system for studying oxidative hemolysis has been used by controling UV-irradiation and concentration of a novel molecular probe, N,N'-bis(2-hydroperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetra-carboxylic diimide (NP-III), which generates hydroxyl radical upon longer wavelength photoirradiation (> 350 nm). NP-III induces 25~30% of hemolysis at low concentration (50 $\mu$M) for 3h-irradiation of UVA. The simultaneous treatment of N-ethylmaleimide (NEM) with NP-IH completely hemotyzed erythrocytes under the same conditions as NP-III alone by both decreasing thiol group and increasing lipid peroxidation in erythrocyte membrane. However. thiol-reducing agents prevented the protein-crosslinking and lipid peroxidation on the NEM-synergistic hemolysis by partially scavenging hydroxyl radical and maintaining the thiol group of erythrocyte membrane in the reduced state. In addition, erythrocytes pretreated with 2,2,5,7,8-pentamethyl-6-hydroxychromane (PMC), vitamin E homologue was able to delay and decrease the lipid peroxidation when compared to cells pretreated with both NEM and PMC. We suggest that the presence of reduced thiols in inner membrane protein by GSH can prevent the protein-crosslinking and the lipid peroxidation, and eventually prevent the oxidative hemolysis of erythrocyte.

• BMB Reports
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• v.29 no.6
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• pp.535-539
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• 1996

The activation of phospholipase D (PLD) catalyzes hydrolysis of phosphatidylcholine (PC) to phosphatidic acid (PA) and choline in plants as well as animals. To determine the presence of PLD in oat cells, we prepared inside-out plasma membrane and cytosolic fractions from oat tissues. PLD activities in both cytosol and plasma membrane were detected by ion chromatography method. The activity of PLD in plasma membrane was dependent upon $Ca^{2+}$ concentration and was heat stable. To investigate whether G-protein couples to PLD, the effects of $GTP{\gamma}S$ and $GDP{\beta}S$ on the PLD activity were measured. PLD activity was dramatically increased 300~400% in the presence of 50 ${\mu}M$ $GTP{\gamma}S$ but not in the presence of 50 ${\mu}M$ $GDP{\beta}S$. These results indicate that G-protein may be involved in regulation of PLD activity. To identify whether PLD is regulated by red light receptor, phytochrome, we irradiated red, far-red, or red/far-red/red light on oat protoplasts. PLD activity has increased 5-fold and 3-fold by treatment with red light and red/far-red/red light, respectively. In contrast, irradiation with far-red light had little or no effect on PLD activity. These results suggest that phytochrome regulates PLD activity through activation of G-protein in oat cells.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• BMB Reports
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• v.39 no.4
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• pp.412-417
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• 2006

We examined the intracellular localization of NS5 protein of Dendrolimus punctatus cytoplasmic polyhedrosis virus (DpCPV) by expressing NS5-GFP fusion protein and proteins from deletion mutants of NS5 in baculovirus recombinant infected insect Spodoptera frugiperda (Sf-9) cells. It was found that the NS5 protein was present at the plasma membrane of the cells, and that the N-terminal portion of the protein played a key role in the localization. A transmembrane region was identified to be present in the N-terminal portion of the protein, and the detailed transmembrane domain (SQIHMVWVKSGLVFF, 57-71aa) of N-terminal portion of NS5 was further determined, which was accorded with the predicted results, these findings suggested that NS5 might have an important function in viral life cycle.

• KSBB Journal
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• v.9 no.3
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• pp.332-338
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• 1994

Solubilization phenomena of a protein in a reversed micellar solution were investigated and a hollow fiber membrane extractor was tested for reversed micellar protein extraction equipment. Alkaline protease was used as a model protein compound and the reversed micellar solution was consisted of AOT and isooctane. It was found that protein solubilization was strongly influenced by ionic strength and pH. The distribution coefficient of the protease between the aqueous solution and the AOT/isooctane solution was also observed to be as high as 4.0 within the scope of this experiment. A hollow fiber membrane extractor was constructed and tested for the protein extraction. The overall mass transfer coefficient at a typical experimental condition of this study was observed to be $6.7{\times}10^{-5}cm/s$. It was also found that the mass transfer resistance on reversed micellar solution was the dominant resistance for the protein transfer.

• Journal of Life Science
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• v.10 no.2
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• pp.131-139
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• 2000

Lecithin-free egg yolk protein (EYP), the by-product of lecithin extraction from egg yolk, which is denatured with an organic solvent, would normally be discarded. In this study, the denatured protein was renatured with alkali, and hydrolyzed with Alcalase in order to utilize by-product. The hydrolysate was separated through a series of ultrafiltration membranes with molecular weight cut-off (MWOO) of 10, 5 and 1 kDa, and the antioxidative activities of the hydrolysates was investigated. The 5K hydrolysate, permeate from 5 kDa membrane, showed stronger antioxidative activity than 10 K and 1 K hydrolysate which were permeated from 10 kDa and 1 kDa membrane, in a linoleic acid autoxidation system. In addition, the optimum concentration of antioxidative activity for 5 K hydrolysate was 1%, and the activity was about 37% higher as compared with α-tocopherol. The synergistic effect was also increased by using the hydrolysates with α-tocopherol.

• Journal of Life Science
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• v.7 no.4
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• pp.322-328
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• 1997

Two phosphorylated proteins having molecular weights of 57kD and 72kD were detected from the slubilized membrane protein fraction of mating type a cells of Rhodosporidium toruloides which belongs to heterobasidiomycetous yeast. The phosphorylation of the protein was inhibited by a sexual pheromone, Rhodotorucine A (Rh. A), which is secreted from mating type a cells. On the other hand, counterpart mating type A cells and M-39 strain which is a styerile mutant derived from a cells, had also the same two phosphorylated proteins, However, the phosphorylation of the protein from A cells, and M-39 strain were not inhibited by the Rh. A. It suggests that inhibition of the phosphorylation reaction by the Rh. A in mating type a cells is a mating-type-specific reaction that relate to transduction mechanism of sexual pheromone signaling.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2001.05a
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• pp.140-140
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• 2001

Porins are major outer membrane proteins which produce non-specific aqueous channels across the membrane that permit the diffusion into the bacterial cells of hydrophilic compounds including sugars, amino acids, and antibiotics. In some gram-negative organisms, antibiotic resistance can be induced by mutational loss of channel that causes a decrease in outer membrane permeability. (omitted)

• Membrane Journal
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• v.31 no.5
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• pp.293-303
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• 2021

Separation membranes used in water filtration, protein purification or biomedical filtration device frequently undergo membrane fouling for several reasons. The formation of biofilm on the membrane surface by bacteria causes a severe problem for durability of the membrane. For the protein separation, the membrane pores get blocked due to surface hydrophobicity of the membrane. There are several approaches controlling the membrane fouling and one of them is the incorporation of silver nanoparticles. Antibacterial properties of silver nanoparticles are well known and thus widely used in several applications. In this review, we have focused on the membranes where silver nanoparticles or its derivatives are either incorporated in the active layer of thin film composite membranes or uniformly distributed throughout the whole membranes.