• Title, Summary, Keyword: 인산화반응

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Methionyl-tRNA-synthetase is a Novel Interacting Protein of LRRK2 (파킨스병 유전인자인 LRRK2와 상호작용하는 methionyl-tRNA synthetase)

  • Kim, Hyejung;Ho, Dong Hwan;Son, Ilhong;Seol, Wongi
    • Journal of Life Science
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    • v.28 no.2
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    • pp.170-175
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    • 2018
  • Parkinson's disease (PD) is the most common movement disorder and the second most common neurodegenerative disease after Alzheimer's disease. Approximately 5~10% of PD patients are familial PD cases. Leucine-rich repeat kinase 2 (LRRK2) has been known to be a causal gene of PD when it is mutated. LRRK2 contains the functional kinase and GTPase domains as well as leucine-rich repeat (LRR) and WD40 domains that are known to play critical roles for protein-protein interaction, suggesting that LRRK2-interacting proteins are important regulators for PD pathogenesis. In an effort to identify proteins interacting with LRRK2, we carried out co-immunoprecipitation of LRRK2 antibody using extracts of NIH3T3 cells that express LRRK2 at a relatively high level. The mass spectrometry analysis of a precipitated band revealed that the co-precipitated band was methionyl-tRNA synthetase (MRS), an ancient enzyme that transfers methionin to its cognate tRNA. The interaction of MRS with LRRK2 was confirmed again by co-immunoprecipitation of endogenous proteins and GST pull-down assays. However, LRRK2 did not phosphorylate recombinant MRS protein in in vitro kinase assays, and over-expression of LRRK2 or MRS did not affect the stability of its partner protein. Our data indicate that LRRK2 interacts with but does not phosphorylate MRS, and the stability of each partner is not affected by the other.

Comparative Study of Nucletic Acid Binding of the Purified RBF Protein and Its Inhibition of PKR phosphorylation (RBF정제단백질의 핵산결합도 및 PKR효소의 인산화억제효과의 비교에 관한 연구)

  • 박희성;김인수
    • Journal of Life Science
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    • v.8 no.2
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    • pp.119-125
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    • 1998
  • Column-purified double-stranded RNA binding factor (RBF) protein was tested for its binding affinity for the different forms of nucleic acids structure such as single-stranded(ss) and double-stranded(ds)RNA and ss- and dsDNA. The RBF protein was incubated with each of these nucleic acid structures in separate reactions and its comparative binding affnity was visualized by SDS-polyacrylamide gel electrophoresis. The RBF protein bound to the dsRNA molecule to form a tight RNA:protein complex in agreement with previous studies, but not to the other nucleic acid molecules confirming its distinctive affinity for the dsRNA structure. In phosphorylation assay in vito, the purified RBF protein significantly inhibited the autophosphorylation of the PKR derived from not only human but mouse source in the presence of poly(I):poly(C). It is suggesting that PKR vs. RBF is similarly under a competitive interaction among different eukaryotic organisms during protein synthesis.

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Effect of Liming on the Extractability of Phosphate Added to Uncultivated Acid Soil (미경지(未耕地)의 산성토양(酸性土壤)에서 시용인산(施用燐酸)의 침출성(浸出性)에 미치는 석회(石灰)의 영향(影響))

  • Yoo, Sun-Ho;Lee, Won-Chu;Park, Sung-O
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.88-94
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    • 1977
  • A laboratory study was made of liming on the extractability of phosphate added to the uncultivated acid soils differing in clay content. The experiments were conducted with different rates of calcium carbonate and triple superphosphate at field capacity and 50% field capacity. The concentration of extractable phosphate was greatly reduced by simultaneous addition of calcium carbonate in the loam soil. When samples of the loam soil were preincubated for ranging from 0 to 50 days in the presence of calcium carbonate prior to phosphate application, the extractable phosphate concentration increased with preincubation time, and more phosphate was extracted from the soil maintained at 50% field capacity during the incubation. However, the extractable phosphate concentration in the sandyloam soil did not change with preincubation time and was not affected by moisture status. It may be concluded that the extractability of added phosphate was influenced by not only liming time but physical properties of soil.

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Production of Fructose 6-Phoschate from Starch Using Thermostable Enzymes (내열성 효소를 이용한 전분으로부터 6-인산과당의 제조)

  • Kwun, Kyu-Hyuk;Cha, Wol-Suk;Kim, Bok-Hee;Shin, Hyun-Jae
    • KSBB Journal
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    • v.22 no.5
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    • pp.345-350
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    • 2007
  • Phosphosugars are found in all living organisms and are commercially valuable compounds with possible applications in the development of a wide range of specialty chemicals and medicines. In carbohydrate metabolism, fructose 6-phosphate (F6P) is an essential intermediate formed by phosphorylation of 6' position of fructose in glycolysis, gluconeogenesis, pentose phosphate pathway and Calvin cycle. In glycolysis, F6P lies within the glycolysis metabolic pathway and is produced by isomerisation of glucose 6-phosphate. For large-scale production, F6P could be produced from starch using many enzymes such as pullulanase, starch phosphorylase, isomerase and mutase. In enzymatic reactions carried out at high temperatures, the solubility of starch is increased and microbial contamination is minimized. Thus, thermophile-derived enzymes are preferred over mesophile-derived enzymes for industrial applications using starch. Recently, we reported the production of glucose 1-phosphate (G1P) from starch by Thermus caldophilus GK24 enzymes. Here we report the production of F6P from starch through three steps; from starch to glucose 1-phosphate (glucan phosphorylase, GP), then glucose 6-phosphate (phosphoglucomutase, GM) and then F6P (phosphoglucoisomerase, GI). Using 200 L of 1.2% soluble starch solution in potassium phosphate buffer, 1,253 g of G1P were produced. Then, 30% yields of F6P were attained at the optimum reaction conditions of GM : G1 (1 : 2.3), 63.5$^{\circ}C$, and pH 6.85. The optimum conditions were found by response surface methodology and the theoretical values were confirmed by the experiments. The optimum starch concentrations were 20 g/L under the given conditions.

Formation of Solution-derived Hydroxyapatite Layer on the Surface of a Shell (용액 반응에 의한 패각 표면의 수산화아파타이트 층 생성 거동)

  • Kim, Hui-Lae;Song, Tae-Woong
    • Journal of the Korean Ceramic Society
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    • v.39 no.12
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    • pp.1177-1182
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    • 2002
  • Highly surfaced and porous hydroxyapatite body was artificially formed on the surface of a shell through a reaction with phosphatic solutions. As a result of qualitative observation, hydroxyapatite seemed to be crystallized by solution-precipitation process accelerated by the nucleation surface of a shell. The process of formation of hydroxyapatite layer was as follows. 1. Dense nucleation and growth on the surface of solid phase 2. Formation of microporous layer by contact and entanglement between crystallines 3. diffusion of solution through the porous layer and thickness growth of layer towards inside

Immobilization of ATP on Bovine $\beta$- Caseins by Using Transglutaminase (효소법에 의한 ATP의 Bovine $\beta$-Casein에의 고정화)

  • 윤세억;박선영김명곤
    • KSBB Journal
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    • v.5 no.3
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    • pp.241-246
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    • 1990
  • ATP analogs were immobilized or bovine caseins by the action of transglutaminase. The ATP analogs immobilized on the caseins were enzymatically active and interconverten by kinases. The immobilized ATP was dephosphorylated to the corresponding ADP by hexokinase and rephosphorylated to the ATP in solid form by acetate kinase. Under the conditions chosen, about 55% of the immobilized ATP was dephosphorylated and about 80% of the resulted ADP was rephosphorylated. Bovine $\beta$-casein was more useful than $\alpha$sf-casein as a carrier and C8-substituted ATP analognwas more effective than N6-substituted one in immobilization. Michaelis constant of C8-substituted ATP analog immobilized on $\beta$-casein was similar to that of free form of ATP and that of ATP analog. The immobilized ATP was much more stable than free ATP and its analog, while maximum velocity was reduced to 37% of the free ATP analog. The immobilized ATP was recovered almost completely by calcium precipitation.

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Design of the Fixed-Bed Catalytic Reactor for the Maleic Anhydride Production (무수마레인산 생산을 위한 고정층 촉매 반응기 설계)

  • Yoon, Young Sam;Koo, Eun Hwa;Park, Pan Wook
    • Applied Chemistry for Engineering
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    • v.10 no.3
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    • pp.467-476
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    • 1999
  • This paper analyzed the behavior of fixed-bed catalytic reactor (FBCR) which synthesizing maleic anhydride(MA) from the selective oxidation of n-butane. The behavior of FBCR describing convection-diffusion-reaction mechanism is examined by using two-dimensional pseudohomogeneous plug-flow transient model, with the kinetics of Langmuir-Hinshelwood type. Prediction model is composed by optimum parameter estimation from temperature profile, yield and conversion of single FBCR on operating condition variations of Sharma's pilot-plant experiment. A double FBCR with same yield and conversion for single FBCR generated a $8.96^{\circ}C$ lower hot spot temperature than a single FBCR. We could predict parametric sensitivity according to the variation of possible operating condition (temperature, concentration, volumetric flow of feed reactant and coolant flow rate) of single and double FBCR. Double FBCR showed the behavior of more operating range than single FBCR. Double FBCR with nonuniform activities could assure safety operation condition for the possible variation of operating condition. Also, double FBCR had slightly higher than the single FBCR in conversion and yield.

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AC Impedance Study of Oxygen Electrode in Phosphoric Acid Fuel Cell (교류 임피던스법에 의한 인산형 연료전지의 산소전극 특성 연구)

  • Song Rak-Hyun;Kim Chang-Soo;Shin Dong-Ryul
    • Journal of the Korean Electrochemical Society
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    • v.3 no.4
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    • pp.191-195
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    • 2000
  • Electrochemical properties of the oxygen electrode in phosphoric acid fuel cell have been studied using AC impedance method as a function of applied potential, operating temperature and teflon content in the electrode. The oxygen electrode reaction in the $105wt.\%$ phosphoric acid is characterized by a parallel resistive component, $R_p$ and a capacitive component, $C_p$ with serial electrolyte resistance, $R_s$. The conductivity of the phosphoric acid is found to be 0.31-0.47 S/cm in the range of $130\;to\;190^{\circ}C$ from the measured impedance. The increase of applied potential and temperature produced the decreased RP and the increased $C_p$, which means the increase of the oxygen electrode reaction rate. The single cells with the cathode of various teflon contents were tested, and the cathode with $40wt.\%$ teflon showed good performance, which is considered to be related to an optimized impedance behavior.

Protein kinase C와 이와 관련된 단백질 연구

  • 이재란;김진한;최명언
    • Proceedings of the Korean Society of Applied Pharmacology
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    • pp.135-135
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    • 1993
  • 신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2+}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

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Treatment of N, P of Auto-Thermal Thermophilic Aerobic Digestion Filtrate with Struvite Crystallization (Struvite 결정화 반응을 이용한 고온 소화 여과액의 N, P 처리 특성)

  • Choo, Yeon-Duk;Kim, Keum-Yong;Ryu, Hong-Duck;Lee, Sang-Ill
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.11
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    • pp.783-789
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    • 2011
  • Recently, auto-thermal thermophilic aerobic digestion (ATAD) has a great attention for destruction of wasted sludge biomass in wastewater treatment plant. Reduction of sludge concentration has been successfully achieved with pilot scale ATAD and ceramic filtration process in field condition. However, high concentration of COD, total nitrogen (TN) and total phosphorus (TP) was observed in filtrate, which should be treated before recirculation of filtrate to biological wastewater treatment plant. This study was focused on removal of nitrogen and phosphorus contained in the filtrate of ATAD, using struvite crystallization method. The effect of operational and environmental parameters (such as, N, P and Mg ion concentration and molar ratio, pH, reaction time, agitation strength, seed dosage, and reaction temperature) on the treatment of TN and TP with struvite crystallization were evaluated. Magnesium (as $MgCl_26H_2O$) and phosphorus (as $K_2HPO_4$) ions were, if necessary, added to increase nitrogen removal efficiency by the crystal formation. Average concentration of $NH_4^+-N$ and $PO_4^{3-}-P$ of the filtrate were 1716.5 mg/L and 325.5 mg/L, respectively. Relationship between removal efficiencies of nitrogen and phosphorus and molar ratios of $Mg^{2+}$ and $PO_4^{3-}-P$ to $NH_4^+-N$ was examined. Crystal formation and nitrogen removal efficiencies were significantly increased as increasing molar ratios of magnesium and phosphorus to nitrogen. As molar ratio of $Mg^{2+}:PO_4^{3-}-P:NH_4^+-N$ were maintained to 2 : 1 : 1 and 2 : 2 : 1, removal efficiencies of nitrogen and phosphorus were 71.6% and 99.9%, and 93.8% and 98.6%, respectively. However, the effect of reaction time, mixing intensity, seed dose and temperature on the struvite crystallization reaction was not significant, comparing to those of molar ratios. Settled sludge volume after struvite crystallization was observed to be reduced with increase of seed dose and to be increased at high temperature.