• Title/Summary/Keyword: $PI3K{\gamma}$

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Cloning and Characterization of Phosphoinositide 3-Kinase γ cDNA from Flounder (Paralichthys olivaceus) (넙치에서 분리된 phosphoinositide 3-kinase γ 유전자의 클로닝 및 특성 연구)

  • Jeong, Tae Hyug;Youn, Joo Yeon;Ji, Keunho;Seo, Yong Bae;Kim, Young Tae
    • Journal of Life Science
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    • v.24 no.4
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    • pp.343-351
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    • 2014
  • Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

Elevated RalA activity in the hippocampus of PI3Kγ knock-out mice lacking NMDAR-dependent long-term depression

  • Sim, Su-Eon;Lee, Hye-Ryeon;Kim, Jae-Ick;Choi, Sun-Lim;Bakes, Joseph;Jang, Deok-Jin;Lee, Kyungmin;Han, Kihoon;Kim, Eunjoon;Kaang, Bong-Kiun
    • BMB Reports
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    • v.46 no.2
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    • pp.103-106
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    • 2013
  • Phosphoinositide 3-kinases (PI3Ks) play key roles in synaptic plasticity and cognitive functions in the brain. We recently found that genetic deletion of $PI3K{\gamma}$, the only known member of class IB PI3Ks, results in impaired N-methyl-D-aspartate receptor-dependent long-term depression (NMDAR-LTD) in the hippocampus. The activity of RalA, a small GTP-binding protein, increases following NMDAR-LTD inducing stimuli, and this increase in RalA activity is essential for inducing NMDAR-LTD. We found that RalA activity increased significantly in $PI3K{\gamma}$ knockout mice. Furthermore, NMDAR-LTD-inducing stimuli did not increase RalA activity in $PI3K{\gamma}$ knockout mice. These results suggest that constitutively increased RalA activity occludes further increases in RalA activity during induction of LTD, causing impaired NMDAR-LTD. We propose that $PI3K{\gamma}$ regulates the activity of RalA, which is one of the molecular mechanisms inducing NMDAR-dependent LTD.

Changes of splenocyte $IFN-{\gamma}$ mRNA synthesis in rats infected with Paragonimus westermani

  • Cho, Jun-Kyong;KWon, Hye-Soo;Joo, Kyoung-Hwan;Lee, Joon-Sang;Cho, Sung-Weon
    • Parasites, Hosts and Diseases
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    • v.37 no.4
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    • pp.285-287
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    • 1999
  • Changes in the expression level of splenocyte $IFN-{\gamma}$mRNA of Sprague-Dawley (SD) rats infected with Paragonimus westermani were analyzed by competitive reverse transcription-polymerase chain reaction (RT-PCR) followed by southern blot. The template RNA was extracted from the splenocytes of rats infected with 20 metacercariae of P. westermani. The products of competitive RT-PCR were subjected to southern blot and enhanced chemiluminescence (ECL), and analyzed with a densitometer. In comparison with that of uninfected control rat splenocytes (value of 1), the levels of mRNA expression of $IFN-{\gamma}$had changed to 0.747 at 1 week post infection (PI), 0.00175 at 2 week PI, 0.0217 at 3 week PI, 0.194 at 4 week PI and then to 0.537 at 5 week PI. The level at 7 week PI had returned to 1.25, comparable with that of uninfected rats. These results show that, when infected with p. westermani, the levels of $IFN-{\gamma}$ mRNA of SD rat splenocytes were remarkably reduced by more than 500 times at 2 week PI and restored to normal level at 7 week PI.

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Thermal and temporal stabilities of a electro-optic coefficient $\gamma_{33}$ in a PI-SOT nonlinear polymer thin film (고분자 박막인 PI-SOT의 전기광학계수 $\gamma_{33}$의 열적 . 시간적 안정성)

  • Jeong, Youn-Hong;Jo, Jae-Heung;Chang, Soo;Kim, Tae;lee, Kwang-Sup
    • Korean Journal of Optics and Photonics
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    • v.10 no.3
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    • pp.188-194
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    • 1999
  • We synthesized the nonlinear optical (NLO) PI-SOT(polyimide system, 4-[N,N-bis(hydroxyethyl)amino-4,-($\beta$-cyano-$\beta$-methylsulfonyl)vinyl]azobenzene) polymer with high electro-optic coefficients as well as good thermal and temporal stabilities of the elector-optic coefficient ${\gamma}_33$ by the simple Mitsunobu reaction. By using the simple reflection method of C. C. Teng, we measured the thermal and temporal stabilities of the electro-optic coefficient ${\gamma}_33$ of corona-poled PI-SOT polymer at the wavelength of 632.8 nm and 852 nm, respectively. At the temperature of $20^{\circ}C$, the electro-optic coefficient ${\gamma}_33$ of corona-poled PI-SOT polymer were 25.12 pm/V at the wavelength of 632.8 nm and 5.40 pm/V at the wavelength of 852 nm. These values were highly stabilized for more than 60 days at 2$0^{\circ}C$ and stabilized within 6% for more than 10 hours at $100^{\circ}C$.

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TILINGS OF PARALLELOGRAMS WITH SIMILAR TRIANGLES

  • Su, Zhanjun;Ding, Ren
    • Journal of applied mathematics & informatics
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    • v.23 no.1_2
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    • pp.321-327
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    • 2007
  • We say that a triangle ${\Delta}$ tiles the polygon ${\rho}\;if\;{\rho}$ can be decomposed into finitely many non-overlapping triangles similar to ${\Delta}$. Let ${\rho}$ be a parallelogram with angles ${\delta}\;and\;{\pi}-{\delta}\;(0<{\delta}{\leq}{\pi}/2)$ and let ${\Delta}$ be a triangle with angles ${\alpha};{\beta},\;{\gamma}\;({\alpha}{\leq}{\beta}{\leq}{\gamma})$. We prove that if ${\Delta}$ tiles ${\rho}$ then either ${\delta}{\in}\;({\alpha},\;{\beta},\;{\gamma},\;{\pi}-{\gamma},\;{\pi}-2{\gamma})\;or\;dimL_{\rho}=dimL_{{\Delta}}$. We also prove that for every parallelogram P, and for every integer n $(where\;n{\geq}2,\;n{\neq}3)$ there is a triangle ${\Delta}$ so that n similar copies of ${\Delta}\;tile\;{\rho}$.

The Heterochromatin-1 Phosphorylation Contributes to TPA-Induced AP-1 Expression

  • Choi, Won Jun
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.308-313
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    • 2014
  • Activator protein-1 (AP-1) is an inducible transcription factor that contributes to the generation of chronic inflammation in response to oxidative and electrophilic stress. Previous studies have demonstrated that the PI3K/Akt1 pathway plays an important role in the transcriptional regulation of AP-1 expression. Although the histone post-translational modifications (PTMs) are assumed to affect the AP-1 transcriptional regulation by the PI3K/Akt pathway, the detailed mechanisms are completely unknown. In the present study, we show that heterochromatin 1 gamma ($HP1{\gamma}$) plays a negative role in TPA-induced c-Jun and c-Fos expression. We show that TPA-induced Akt1 directly phosphorylates $HP1{\gamma}$, abrogates its suppressive function and increases the interaction between histone H3 and 14-3-$3{\varepsilon}$. Collectively, these our data illustrate that the activation of PI3K/Akt pathway may play a permissive role in the recruitment of histone readers or other coactivators on the chromatin, thereby affecting the degree of AP-1 transcription.

Development of screening systems for modulators on phospholipase-mediated signal transduction

  • Lee, Young-Han-;Min, Do-Sik;Kim, Jae-Ho-;Suh, Pann-Ghill;Ryu, Sung-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.186-186
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    • 1994
  • Many agonists have been known to activate the hydrolysis of membrane phospholipids through the bindings with corresponding receptors on the various cells. Diacylglycerol and inositol 1,4,5-trisphosphate(IP3) generated by the action of phosphoinositide-specific phospholipase C (PI-PLC) are well known second messengers for the activation of protein kinase C and the mobilization of Ca2+ in many cells. Three types of PI-PLC isozyme (${\alpha}$,${\gamma}$, and $\delta$) and several subtrpes for each type have been identified from mammalian sources by purification of enzymes and cloning of their cDNAs. Each type PI-PLC isozyme is coupled to different receptors and mediators, for example, ${\beta}$-types are coupled to the seven-transmembrane-receptors via Gq family of G-proteins and ${\beta}$-types directly to the receptor tyrosine kinases. Specific modulators for the signaling pathway through each type of PI-PLC should be very useful as potential potential candidates for lend substances in developing novel drugs. To establish the sensitive and convenient screening systems for searching modulators on PI-PLC mediated signaling, two kinds of approaches have been tried. (1) Establishment of in vitro assay condition for each type of PI-PLC isozyme: Overexpression by using vaccinia virus and purification of each isozyme was carried out for the preparation of large amounts of enaymes. Optimum and sensitive assay condition for the measurements of PI-ELC activities were established. (2) Development of the cell lines in which each type of PI-PLC is permanently overexpressed: A fibroblast cell line (3T3${\gamma}$1-7) in which PI-PLC-${\gamma}$1 was overexpressed by using pZip-neo expression vector was developed and used for the measurement of PDGF-induced IP3 formation. The responses for IP3 formed in 3T3${\gamma}$1-7 cells by the treatment of PDGF is 8 times more sensitive than those in control cells. 3T3${\gamma}$l-7 cell is useful for the screening of the inhibitors on the PDGF-induced cellular responses from large number of samples in a small volume(50 ${\mu}$l) and short time(5-15 min). Using these systems, we screened hundreds of herb-extracts for the inhibition of PDGF-induced IP3 formation and selected several extracts that showed the inhibition as the candidates for isolation and characterization of active substances. The determination of the acting point of selected extracts or fractions in the PDGF signaling pathway has been analyzing.

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REGULARITY OF SOAP FILM-LIKE SURFACES SPANNING GRAPHS IN A RIEMANNIAN MANIFOLD

  • Gulliver, Robert;Park, Sung-Ho;Pyo, Jun-Cheol;Seo, Keom-Kyo
    • Journal of the Korean Mathematical Society
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    • v.47 no.5
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    • pp.967-983
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    • 2010
  • Let M be an n-dimensional complete simply connected Riemannian manifold with sectional curvature bounded above by a nonpositive constant $-{\kappa}^2$. Using the cone total curvature TC($\Gamma$) of a graph $\Gamma$ which was introduced by Gulliver and Yamada [8], we prove that the density at any point of a soap film-like surface $\Sigma$ spanning a graph $\Gamma\;\subset\;M$ is less than or equal to $\frac{1}{2\pi}\{TC(\Gamma)-{\kappa}^2Area(p{\times}\Gamma)\}$. From this density estimate we obtain the regularity theorems for soap film-like surfaces spanning graphs with small total curvature. In particular, when n = 3, this density estimate implies that if $TC(\Gamma)$ < $3.649{\pi}\;+\;{\kappa}^2\inf\limits_{p{\in}F}Area(p{\times}{\Gamma})$, then the only possible singularities of a piecewise smooth (M, 0, $\delta$)-minimizing set $\Sigma$ are the Y-singularity cone. In a manifold with sectional curvature bounded above by $b^2$ and diameter bounded by $\pi$/b, we obtain similar results for any soap film-like surfaces spanning a graph with the corresponding bound on cone total curvature.

T cell phenotype and intracellular $IFN-{\gamma}$ production in peritoneal exudate cells and gut intraepithelial lymphocytes during acute Toxoplasma gondii infection in mice

  • Lee, Young-Ha;Shin, Dae-Whan
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.119-129
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    • 2002
  • Although there are many reports on the splenic (systemic) T cell response after Toxoptasma gondii infection, little information is available regarding the local T cell responses of peritoneal exudate cells (PEC) and gut intraepithelial Iymphocytes (IEL) following peroral infection with bradyzoites. Mice were infected with 40 cysts of the 76K strain of T. gondii, and then sacrificed at days 0, 1, 4, 7 and 10 postinfection (PI). The cellular composition and T cell responses of PEC and IEL were analyzed. The total number of PEC and IEL per mouse increased after infection, but the ratio of increase was higher in IEL. Lymphocytes were the major component of both PEC and IEL. The relative percentages of PEC macrophages and neutrophils/eosinophils increased signiflcantly at day 1 and 4 PI, whereas those of IEL did not change significantly. The percentage of PEC NK1.1 and ${\gamma\delta}T$ cells peaked at day 4 PI (p < 0.0001), and CD4 and $CD8{\alpha}T$ cells increased continuously after infection. The percentages of IEL $CD8{\alpha}$ and ${\gamma\delta}T$ cells decreased slightly at first, and then increased. CD4 and NK1.1 T cells of IEL did not change significantly after infection. $IFN-{\gamma}-producing$ PEC NK1.1 T cells increased significantly from day 1 PI, but the other T cell subsets produced $IFN-{\gamma}$ abundantly thereafter. The proportion of IEL $IFN-{\gamma}-producing$ $CD8{\alpha}$ and ${\gamma\delta}T$ cells increased significantly after infection, while IEL NK1.1 T cells had similar $IFN-{\gamma}$ production patterns. Taken together, CD4 T cells were the major phenotype and the important $IFN-{\gamma}$ producing T cell subsets in PEC after oral infection with T. gondii whereas $CD8{\alpha}T$ cells had these roles in IEL. These results suggest that PEC and IEL comprise different cell differentials and T cell responses, and according to infection route these factors may contribute to the different cellular immune responses.