• Title/Summary/Keyword: GTP-Binding Protein

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Regulation of c-fos promoter through interaction between dopamine D3 receptor and RGL, ral GDP dissociation stimulator-like

  • Park, Ju-Ran;Kim, So-Young;Kim, Kyeong-Man
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.89.2-89.2
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    • 2003
  • Ral GDP dissociation stimulator (Ral GDS) has been found to be an effector protein of Ras, and Ral, a member of small GTP-binding protein (G protein) superfamily, has been suggested to act downstream of Ras. Ral GDP dissociation stimulator-like (RGL) shares 50% amino acid identity with Ral GDP dissociation stimulator, and assumed to possess similar functional role. Using yeast two-hybrid screening, we found that dopamine D3 receptor interacts with RGL. Since RGL is known to regulate the expression of c-fos promoter, effects of D3R on gene expression of c-fos promoter was studied. (omitted)

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Identification of Three Positive Regulators in the Geldanamycin PKS Gene Cluster of Streptomyces hygroscopicus JCM4427

  • Kim, Won-Cheol;Lee, Jung-Joon;Paik, Sang-Gi;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1484-1490
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    • 2010
  • In the Streptomyces hygroscopicus JCM4427 geldanamycin biosynthetic gene cluster, five putative regulatory genes were identified by protein homology searching. Among those genes, gel14, gel17, and gel19 are located downstream of polyketide synthase genes. Gel14 and Gel17 are members of the LAL family of transcriptional regulators, including an ATP/GTP-binding domain at the N-terminus and a DNA-binding helix-turn-helix domain at the C-terminus. Gel19 is a member of the TetR family of transcriptional regulators, which generally act to repress transcription. To verify the biological significance of the putative regulators in geldanamycin production, they were individually characterized by gene disruption, genetic complementation, and transcriptional analyses. All three genes were confirmed as positive regulators of geldanamycin production. Specifically, Gel17 and Gel19 are required for gel14 as well as gelA gene expression.

Affinity between TBC1D4 (AS160) phosphotyrosine-binding domain and insulin-regulated aminopeptidase cytoplasmic domain measured by isothermal titration calorimetry

  • Park, Sang-Youn;Kim, Keon-Young;Kim, Sun-Min;Yu, Young-Seok
    • BMB Reports
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    • v.45 no.6
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    • pp.360-364
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    • 2012
  • Uptake of circulating glucose into the cells happens via the insulin-mediated signalling pathway, which translocates the glucose transporter 4 (GLUT4) vesicles from the intracellular compartment to the plasma membrane. Rab GTPases are involved in this vesicle trafficking, where Rab GTPases-activating proteins (RabGAP) enhance the GTP to GDP hydrolysis. TBC1D4 (AS160) and TBC1D1 are functional RabGAPs in the adipocytes and the skeletonal myocytes, respectively. These proteins contain two phosphotyrosine-binding domains (PTBs) at the amino-terminus of the catalytic RabGAP domain. The second PTB has been shown to interact with the cytoplasmic region of the insulin-regulated aminopeptidase (IRAP) of the GLUT4 vesicle. In this study, we quantitatively measured the ${\sim}{\mu}M$ affinity ($K_D$) between TBC1D4 PTB and IRAP using isothermal titration calorimetry, and further showed that IRAP residues 1-49 are the major region mediating this interaction. We also demonstrated that the IRAP residues 1-15 are necessary but not sufficient for the PTB interaction.

Regulatgion of the Transport of Vitellogenin by Heterotrimeric G-Proteins during Oogenesis of a Polychaete, Pseudopotamilla occelata

  • Yi, Bong-Kyung;Lee, Yang-Rim
    • Animal cells and systems
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    • v.2 no.1
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    • pp.93-97
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    • 1998
  • Coelomoic fluid protein (CP), a vitellogenin contained in the coelomic fluid of polychaetes, is transported by receptor-mediated endocvtosis that is controlled by GTP-binding proteins. Transport of 125l-CP was markedly inhibited by AlF4 and toxins such as cholera toxin and pertussis toxin. These effects appear to be mediated by cAMP, since 125l-CP transport was also greatly inhibited by dibutyryl cAMP. The results strongly suggest that hetero trimeric G-protein is involved in the regulation of 125l=CP transport through the activation of adenylyl cyclase. Immunoblotting tests with antibodies against Gsa and Gia subunits showed a Gsa subunit of 45 kDa in the membrane of oocytes of intermediate and large size classes and a Gia subunit of 41 kDa only in the oocytes of the intermediate size class.

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γ-Glutamyltranspeptidase Gene from Bacillus subtilis BS 62 (Bacillus subtilis BS 62의 γ-Glutamyltranspeptidase 유전자)

  • Lee, Tae-Eun;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.34 no.2
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    • pp.161-170
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    • 2007
  • To characterize $\gamma$-glutamyltranspeptidase ($\gamma$-GTP or ggt; EC 2. 3. 2. 2.) gene of Bacillus subtilis BS 62, the $\gamma$-GTP gene of BS 62 was prepared from PCR products amplified with the chromosomal DNA. The $\gamma$-GTP gene of about 2.5 kb was sequenced, and its homology was compared with the other ggt genes which were reported previously. The base sequence of the gene appeared to have an open reading frame of 1,758 bp encoding a protein of 62,175 Da. The coding region was flanked by putative ribosome binding site - AGGAGG of 7th to 12th upstream - and the stem-loof sequence was followed by transcription terminator codon. Homology of the amino acid residues sequence consisting of 587 amino acid residues was found as 98% with Bacillus subtilis gene (BSU49358), 97.4% with that of Bacillus subtilis KX 102, 37% with Pseudomonas sp. A14 (S63255) and 38% with Streptomyces avermitils (AP005028).

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Atypical formations of gintonin lysophosphatidic acids as new materials and their beneficial effects on degenerative diseases

  • Ji-Hun Kim;Ra Mi Lee;Hyo-Bin Oh;Tae-Young Kim;Hyewhon Rhim;Yoon Kyung Choi;Jong-Hoon Kim;Seikwan Oh;Do-Geun Kim;Ik-Hyun Cho;Seung-Yeol Nah
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.1-11
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    • 2024
  • Fresh ginseng is prone to spoilage due to its high moisture content. For long-term storage, most fresh ginsengs are dried to white ginseng (WG) or steamed for hours at high temperature/pressure and dried to form Korean Red ginseng (KRG). They are further processed for ginseng products when subjected to hot water extraction/concentration under pressure. These WG or KRG preparation processes affect ginsenoside compositions and also other ginseng components, probably during treatments like steaming and drying, to form diverse bioactive phospholipids. It is known that ginseng contains high amounts of gintonin lysophosphatidic acids (LPAs). LPAs are simple lipid-derived growth factors in animals and humans and act as exogenous ligands of six GTP-binding-protein coupled LPA receptor subtypes. LPAs play diverse roles ranging from brain development to hair growth in animals and humans. LPA-mediated signaling pathways involve various GTP-binding proteins to regulate downstream pathways like [Ca2+]i transient induction. Recent studies have shown that gintonin exhibits anti-Alzheimer's disease and antiarthritis effects in vitro and in vivo mediated by gintonin LPAs, the active ingredients of gintonin, a ginseng-derived neurotrophin. However, little is known about how gintonin LPAs are formed in high amounts in ginseng compared to other herbs. This review introduces atypical or non-enzymatic pathways under the conversion of ginseng phospholipids into gintonin LPAs during steaming and extraction/concentration processes, which exert beneficial effects against degenerative diseases, including Alzheimer's disease and arthritis in animals and humans via LPA receptors.

Possible target for G protein antagonist: Identification of specific amino acid residue responsible for the molecular interaction of G$\alpha$ 16 with chemoattractant C5a receptor.

  • 이창호
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2000.04a
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    • pp.17-19
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    • 2000
  • Heterotrimeric G Proteins transduce ligand binding to a wide variety of seven transmembrane cell surface receptors into intracellular signals. The currently accepted model for the activation of G protein suggests that ligand-activated receptor accelerates GDP-GTP exchange reactions on the ${\alpha}$ subunit of the heterotrimeric G protein. At least seventeen distinct isoforms of the G${\alpha}$ subunit protein have been identified in mammalian organisms. Among them, the G${\alpha}$q family consists of five members whose ${\alpha}$ subunits show different expression patterns. G${\alpha}$q and G${\alpha}$11 seem to be almost ubiquitously expressed, whereas G${\alpha}$14 is predominantly expressed in spleen, lung, kidney and testis. G${\alpha}$16 and its murine counterpart G${\alpha}$15 are expressed in hematopoietic cells and has been shown to couple a wide variety of receptors to phosphoinositide-specific phospholipase C activity. Beta-isoforms of phospholipase C were shown to be activated by all members of G${\alpha}$q family, i.e., G${\alpha}$q, G${\alpha}$11, G${\alpha}$l4 and G${\alpha}$16 subunits either in reconstitution system. or in experiments using cDNA transfection with intact Cos-7 cells.

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Expression of a Heat Shock Protein 70 (Hsp70) in Red Seabream Pagrus major Infected with Longicollum pagrosomi (구두충(Longicollum pagrosomi)에 감염된 참돔(Pagrus major)의 Heat Shock Protein 70 (Hsp70) 발현)

  • Park, Hyung-Jun;Min, Byung-Hwa
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.2
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    • pp.163-169
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    • 2018
  • This study examined the expression of heat shock protein 70 (Hsp70) in red seabream Pagrus major infected by the, acanthocephalan parasites Longicollum pagrosomi. We cloned the full-length Hsp70 cDNA from the liver of the red seabream. The full-length cDNA had a 1,950 bp open reading frame (ORF) that encoded a protein of 650 amino acids. The deduced amino acid sequence of Hsp70 contained all of the conserved Hsp70 family signature sequences and an adenosine triphosphate (ATP)/guanosine triphosphate (GTP) binding motif, including the EEVD (consensus sequence that terminates in Hsp70 family) consensus sequence. The expression of Hsp70 mRNA was upregulated int the fish head-kidney and liver, as determined by quantitative real-time PCR. We quantified the Hsp70 mRNA expression in normal red seabream and fish infected fish by L. pagrosomi. The expression of Hsp70 mRNA was significantly higher in the infected red seabream. These results suggest that Hsp70 play a role of protection against stress and inflammation caused by the parasite and may help maintain homeostasis.

Relationship of the Signal Transduction-mediated Proteins and Enzymes to Contractility and Plasticity in Skeletal Muscles (골격근의 수축과 가소성에 대한 신호전달-매개 단백질 및 관련 효소의 상관성)

  • Kim, Jung-Hwan
    • The Journal of Korean Physical Therapy
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    • v.19 no.4
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    • pp.1-14
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    • 2007
  • Background: It is generally accepted that skeletal muscle contraction is triggered by nerve impulse and intracellular $Ca^{2+}\;([Ca^{2+}]_i)$ released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR). Specifically, this process, called excitation-contraction (E-C) coupling, takes place at intracellular junctions between the plasma membrane, the transverse (T) tubule L-type $Ca^{2+}$ channel (dihydropyridine-sensitive L-rype $Ca^{2+}$ channel, DHPR, also called tetrads), and the SR $Ca^{2+}$ release channel (ryanodine-sensitive $Ca^{2+}$ release channel, RyR, also called feet) of internal $Ca^{2+}$ stores in skeletal muscle cells. Furthermore, it has been reported that the $Ca^{2+-}$ dependent and -independent contraction determine the expression of skeletal muscle genes, thus providing a mechanism for tightly coupling the extent of muscle contraction to regulation of muscle plasticity-related excitation-transcription (E-T) coupling. Purpose: Expression and activity of plasticity-associated enzymes in gastrocnemius muscle strips have not been well studied, however. Methods: Therefore, in this study the expression and phosphorylation of E-C and E-T coupling-related mediators such as protein kinases, ROS(reactive oxygen species)- and apoptosis-related substances, and others in gastrocnemius muscles from rats was examined. Results: I found that expression and activity of MAPKs (mitogen-activated protein kinases, ERK1/2, p38MAPK, and SAPK/JNK), apoptotic proteins (cleaved caspase-3, cytochrome c, Ref-1, Bad), small GTP-binding proteins (RhoA and Cdc42), actin-binding protein (cofilin), PKC (protein kinase C) and $Ca^{2+}$ channel (transient receptor potential channel 6, TRPC6) was observed in rat gastrocnemius muscle strips. Conclusion: These results suggest that MAPKs, ROS- and apoptosis-related enzymes, cytoskeleton-regulated proteins, and $Ca^{2+}$ channel may in part functionally import in E-C and E-T coupling from rat skeletal muscles.

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Rhodopsin Chromophore Formation and Thermal Stabilities in the Opsin Mutant E134Q/M257Y (옵신 mutant E134Q/M257Y의 로돕신 형성과 열안정성 분석)

  • Kim, Jong-Myoung
    • Journal of Life Science
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    • v.22 no.7
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    • pp.863-870
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    • 2012
  • Rhodopsin, a dim light photoreceptor, has been regarded as one of the model systems for the structural and functional study of G protein-coupled receptors (GPCRs). Constitutively active mutant GPCRs leading to the activation of heterotrimeric GDP/GTP-binding protein signaling in the absence of ligand binding are of interest for the study of the activation mechanism in GPCRs. The present study focused on the opsin mutant E134Q/M257Y, which showed a moderate level of constitutive activity and the formation of two distinct rhodopsin chromophores with absorption maxima of 500 nm and 380 nm, depending on the presence of an inverse agonist, 11-cis-retinal, and an agonist, all-trans-retinal, respectively. Reconstitution of the mutant rhodopsin upon incubation with different ratios of 11-cis-retinal and the all-trans-retinal, as well as upon sequential binding of the two retinals, indicated its preferential binding to 11-cis-retinal. The thermal stability of the 11-cis-retinal-bound form of the E134Q/M257Y mutant was lower than that of the mutants containing a single replacement but higher than that of the all-trans-retinal-bound forms. The mutant also showed a lower stability in its opsin state as compared with that of the wild-type opsin but had little effects on the binding affinity to 11-cis-retinal. Information obtained in this study will be helpful for analyzing the structural changes associated with the activation of rhodopsin and GPCRs.