• Title/Summary/Keyword: ser-M

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The Purification and Characterization of Bacillus subtilis Tripeptidase (PepT)

  • Park, Yong-Seek;Cha, Myung-Hoon;Yong, Whan-Mi;Kim, Hyo-Joon;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • v.32 no.3
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    • pp.239-246
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    • 1999
  • A tripeptidase (PepT) was purified to homogeneity from Bacillus subtilis through four sequential chromatographies including DEAE-Sepharose ion exchange, hydroxylapatite, mono-Q FPLC ion exchange, and Superose-12 FPLC gel filtration. The apparent molecular mass of the enzyme was 49,200 Da and 51,400 Da as determined by sodium dodecylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, and the enzyme exists in a monomeric form. The physicochemical properties of the enzyme were as follows: optimum pH at 7.5, optimum temperature at $60^{\circ}C$, and pI at 4.9. The $K_m$ and $V_{max}$ values of the enzyme were 4.3 mM and 2.5 mmol/min/mg, respectively, with MetAla-Ser as substrate. The B. subtilis PepT requires $Co^{2+}$ ion(s) for activation, while it is inactivated by EOTA and 1,10-phenanthroline, suggesting that it is a metalloprotein. The enzyme was not inhibited by any of serine protease, aspartic protease, or leucine aminopeptidase inhibitors. The enzyme showed comparable activities towards four different substrates including Met-Ala-Ser, Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The amino terminal sequence of PepT determined by Edman degradation was found to be MKEEIIERFTTYVXV and turned out to be identical to that of PepT deduced from a cloned B. subtilis pepT.

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Ameliorative Effects of Ombuoside on Dopamine Biosynthesis in PC12 Cells

  • Davaasambuu, Uchralsaikhan;Park, Keun Hong;Park, Hyun Jin;Choi, Hyun Sook;Lee, Chong Kil;Hwang, Bang Yeon;Lee, Myung Koo
    • Natural Product Sciences
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    • v.24 no.2
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    • pp.99-102
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    • 2018
  • This study investigated the effects of ombuoside, a flavonol glycoside, on dopamine biosynthesis in PC12 cells. Ombuoside at concentrations of 1, 5, and $10{\mu}M$ increased intracellular dopamine levels at 1 - 24 h. Ombuoside (1, 5, and $10{\mu}M$) also significantly increased the phosphorylation of tyrosine hydroxylase (TH) (Ser40) and cyclic AMP-response element binding protein (CREB) (Ser133) at 0.5 - 6 h. In addition, ombuoside (1, 5, and $10{\mu}M$) combined with L-DOPA (20, 100, and $200{\mu}M$) further increased intracellular dopamine levels for 24 h compared to L-DOPA alone. These results suggest that ombuoside regulates dopamine biosynthesis by modulating TH and CREB activation in PC12 cells.

A Performance Comparison of mSE-MMA and mDSE-MMA Adaptive Equalization Algorithm in 16-QAM Signal Transmission (16-QAM 신호 전송에서 mSE-MMA와 mDSE-MMA 적응 등화 알고리즘의 성능 비교)

  • Lim, Seung-Gag
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.21 no.2
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    • pp.61-66
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    • 2021
  • This paper related with the performance comparison of mSE-MMA and mDSE-MMA adaptive equalization algorithm which is possible to reduce the intersymbol interference that occurs in the nonlinear communication channel transmitting 16-QAM signal. This two algorithm is possible to reduce the computational load compared to the current MMA algorithm, it has the degraded equalization performance due to simplified arithmetic in order to applying the mobile communication terminal. In order to improve the performance degradation, they controls the step size according to the existence of arbitrary radius circle of equalizer output compared to transmitted symbol point. The variation of step size according to this principle is applied to the SE-MMA and DSE-MMA, namely mSE-MMA and mDSE-MMA algorithm, the algorithm's performance were compared in the same channel and noise environment by computer simulation. As a result of simulation, the mSE-MMA has more superior to the mDSE-MMA in residual value of every performance index and SER performance, and the vice versa result in convergence speed.

Purification and Characterization of the Functional Catalytic Domain of PKR-Like Endoplasmic Reticulum Kinase Expressed in Escherichia coli

  • Yun Jin-A;Chung Ho-Young;Kim Seong-Jun;Cho Hyun-Soo;Oh Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1453-1458
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    • 2006
  • PKR-like endoplasmic reticulum (ER) kinase (PERK) is a type I transmembrane ER-resident protein containing a cytoplasmic catalytic domain with a Ser/Thr kinase activity, which is most closely related to the eukaryotic translation initiation factor-$2{\alpha}$ ($eIF2{\alpha}$) kinase PKR involved in the antiviral defense pathway by interferon. We cloned and expressed the PERK C-terminal kinase domain (cPERK) in Escherichia coli. Like PERK activation in cells under ER stress, wild-type cPERK underwent autophosphorylation when overexpressed in E. coli, whereas the cPERK(K621M) with a methionine substitution for the lysine at amino acid 621 lost the autophosphorylation activity. The activated form cPERK which was purified to near homogeneity, formed an oligomer and was able to trans-phosphorylate specifically its cellular substrate $eIF2{\alpha}$. Two-dimensional phosphoamino acids analysis revealed that phosphorylation of cPERK occurs at the Ser and Thr residues. The functionally active recombinant cPERK, and its inactive mutant should be useful for the analysis of biochemical functions of PERK and for the determination of their three-dimensional structures.

Performance Evaluation of iSER on Storage system using Infiniband fabric (인피니밴드기반 저장장치에서의 iSER(iSCSI Extension for RDMA) 성능평가)

  • Kim, Young Hwan;Son, Jae-Gi;Jung, Hye-Dong
    • Proceedings of the Korea Information Processing Society Conference
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    • 2013.05a
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    • pp.148-150
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    • 2013
  • 최근 TCP/IP에서 세션을 통하여 노드들 간의 통신을 연결하는 방식에서 현재는 하나의 채널을 통해 고속의 I/O가 가능하도록 하는 인피니밴드 같은 기술이 많이 연구되고 있다. 인피니밴드는 프로세싱 노드와 입출력 장치 사이의 통신, 프로세스간 통신에 대한 산업 표준이 되고 있고 프로세싱 노드와 입출력 장치를 연결하기 위해 스위치 기반의 상호 연결은 전통적인 버스 입출력을 대체하는 새로운 입출력 방식이다. 또한 인피니밴드에서는 현재 이슈가 되고 있는 RDMA 방식을 이용해 원격지 서버들간에 직접 메모리 접근 방식을 통해 CPU와 OS의 로드를 최소화하고 있다. 본 논문에서는 인피니밴드 네트워크를 이용하는 저장장치 접근 프로토콜인 iSER(iSCSI Extension RDMA Protocol)와 기존 이더넷망에서 사용되는 iSCSI(Internet SCSI) 프로토콜을 이용하여 서버와 저장장치 간의 IOPS 와 초당 데이터 전송량에 대한 성능을 평가한다. 우리는 성능평가를 위해 Intel에서 제공하는 저장장치 I/O 성능평가 도구인 IO meter를 이용했다.

Generation of Nucler Hybrids Overcoming the Natural Barrier of Incompatibility: Transfer of Nuclei from Lentinula edodes into Protoplasts of Coriolus versicolor

  • Kim, Chaekyun;Choi, Eung-Chil;Kim, Byoung-Kak
    • Archives of Pharmacal Research
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    • v.23 no.1
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    • pp.79-86
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    • 2000
  • Heterokaryotic nuclear hybrids overcoming the natural barriers of incompatibility have been studied in basidiomycetes. To produce these nuclear hybrids between incompatible mushrooms, which have several potent pharmacological effects, nuclear transfer was performed between Lentinula edodes and Coriolus versicolor. Nuclei from serine auxotrophs of Lentinula edodes, $LE207(Ser^{-})$ were transferred into the protoplasts of arginine auxotrophs of Coriolus versicolor, $CV17(Ser^{-})$using 30% polyethylene glycol 4000 in 10 mM $Cacl_{2}$-glycine solution (pH 8.0). Nulcear transfer progenies were selected by nutritional complementation on minimal media supplemented with 0.6 M sucrose. The progenies were classified based on colony morphology to L. edodes-like, C, versicolor-like and non-parental type. Most of the progenies grew slower than either parent. The number of nuclei per cell was similar but the DNA content varied between progenies. The isozyme patterns of nuclear hybrids resembled either of the parent porfiles or showed a mixed profile.

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Dephosphorylation of p53 Ser 392 Enhances Trimethylation of Histone H3 Lys 9 via SUV39h1 Stabilization in CK2 Downregulation-Mediated Senescence

  • Park, Jeong-Woo;Bae, Young-Seuk
    • Molecules and Cells
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    • v.42 no.11
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    • pp.773-782
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    • 2019
  • Cellular senescence is an irreversible form of cell cycle arrest. Senescent cells have a unique gene expression profile that is frequently accompanied by senescence-associated heterochromatic foci (SAHFs). Protein kinase CK2 (CK2) downregulation can induce trimethylation of histone H3 Lys 9 (H3K9me3) and SAHFs formation by activating SUV39h1. Here, we present evidence that the PI3K-AKT-mTOR-reactive oxygen species-p53 pathway is necessary for CK2 downregulation-mediated H3K9me3 and SAHFs formation. CK2 downregulation promotes SUV39h1 stability by inhibiting its proteasomal degradation in a p53-dependent manner. Moreover, the dephosphorylation status of Ser 392 on p53, a possible CK2 target site, enhances the nuclear import and subsequent stabilization of SUV39h1 by inhibiting the interactions between p53, MDM2, and SUV39h1. Furthermore, $p21^{Cip1/WAF1}$ is required for CK2 downregulation-mediated H3K9me3, and dephosphorylation of Ser 392 on p53 is important for efficient transcription of $p21^{Cip1/WAF}$. Taken together, these results suggest that CK2 downregulation induces dephosphorylation of Ser 392 on p53, which subsequently increases the stability of SUV39h1 and the expression of $p21^{Cip1/WAF1}$, leading to H3K9me3 and SAHFs formation.

A Performance Evaluation of mDSE-MMA Adaptive Equalization Algorithm in QAM Signal (QAM 신호에서 mDSE-MMA 적응 등화 알고리즘의 성능 평가)

  • Lim, Seung-Gag
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.20 no.2
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    • pp.103-108
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    • 2020
  • This paper related with the performance evaluation of mDSE-MMA adaptive equalization algorithm which is possible to reduce the distortion that occurs in nonlinear communication channel like as additive noise, intersymbol interference and fading when transmitting the QAM signal. The DSE-MMA algorithm is possible to reduce the computational load compared to the presently MMA algorithm, it has the degraded equalization performance by this. In order to improve the performance degradation of DSE-MMA, the mDSE-MMA controls the step size according to the existence of arbitrary radius circle of equalizer output is centered at transmitted symbol point. The performance of proposed mDSE-MMA algorithm were compared to present DSE-MMA using the same channel and noise environment by computer simulation. For this, the recoverd signal constellation which is the output of equalizer, residual isi and MD, MSE learning curve which is represents the convergence performance and SER were applied as performance index. As a result of simulation, the mDSE-MMA has more superior to the DSE-MMA in every performance index.

Influence of Energy Restriction and Pre-incubation Holding Period of Eggs on Fertility and Hatchability in Aged Broiler Breeders

  • Shyam Sunder, G.;Vijaya Kumar, Ch.;Panda, A.K.;Rama Rao, S.V.;Raju, M.V.L.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.240-245
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    • 2010
  • The effects of controlled energy restriction and duration of pre-incubation egg holding on fertility, hatchability and hatch losses were evaluated in aged broiler breeders (64 wk). The energy (ME) required for maintenance, activity, growth and anticipated egg production was calculated and offered to a control group (283-471 kcal/kg) from 21-64 weeks of age. In three other groups, ME was quantitatively reduced either by 20% (SER; severe energy restriction) or 10% (MER; moderate energy restriction) and increased by10% (EEF; excess energy feeding) over the control group (CER; controlled energy restriction). Each diet was offered to 130 pullets in individual cages, and the quantity of ME increased with age. At the end of 64 weeks, fertile eggs were collected from each dietary group for 11 consecutive days and grouped under 4 holding periods based on the length of storage (2, 5, 8 or 11 d). The influence of energy regimes, egg holding intervals and their interaction was evaluated on fertility, hatch losses and hatchability. Broiler breeders maintained on SER regime (231-419 kcal/d) produced maximum number of eggs (993) followed by MER (819), CER (624) and EEF (438) during the 11-day period. The percent fertility and hatchability was significantly (p$\leq$0.05) higher in SER and MER groups compared to CER and EEF. However, energy regimes did not influence the loss in egg weight during pre-incubation storage, shell weight, shell thickness or hatch losses as dead germs and dead in shell. The improvement in hatchability in SER and MER groups appeared to be closely related to higher fertility and lower embryonic mortality. Holding of eggs for 11 days showed a linear loss in egg weight with the length of storage, but did not influence the fertility and hatch losses. The percent hatchability on eggs set was maximum when storage period was restricted to 5 days. The interaction between energy regimes and egg holding periods exhibited better hatchability results with SER regime when eggs were held for 5 days. Response to MER was not different from SER. It was obvious that energy restriction during production period had a positive influence on egg number, fertility and hatchability in aged breeders. At 64 weeks of age, holding of fertile eggs for 5 days prior to incubation was adequate for optimum hatchability in breeders.

Identification and Characterization of a Novel Antioxidant Peptide from Bovine Skim Milk Fermented by Lactococcus lactis SL6

  • Kim, Sang Hoon;Lee, Ji Yoon;Balolong, Marilen P.;Kim, Jin-Eung;Paik, Hyun-Dong;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.37 no.3
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    • pp.402-409
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    • 2017
  • A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.