• 제목/요약/키워드: subunits$(\alpha'-,\

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A New Synthesis of Triphenylphosphorane Ylide Precursors to α-Keto Amide/Ester and Tricarbonyl Units via Horner-Wadsworth-Emmons Reaction

  • Lee, Kie-Seung
    • Bulletin of the Korean Chemical Society
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    • 제31권10호
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    • pp.2776-2782
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    • 2010
  • Newly developed Horner-Wadsworth-Emmons (HWE) reagents 5 having triphenylphosphorane ylide subunits readily condensed with various carbonyl compounds under mild reaction conditions to afford $\beta,\gamma$-unsaturated $\alpha$-keto triphenylphorane ylides in good to excellent yields, which were hydrogenated over Pd-C (10%)/$H_2$ (1 atm) to give the corresponding $\alpha$-keto triphenylphorane ylides in quasi-quantitative yields. These triphenyphosphorane ylides have been utilized as the precursors to $\alpha$-keto amide/ester and vicinal tricarbonyl units in Wasserman's synthetic protocols, and have previously been prepared only from carboxylic acids/acid chlorides. Our new approaches provide excellent alternatives for the synthesis of triphenylphosphorane ylide precursors to $\alpha$-keto amide/ester and vicinal tricarbonyl units directly from carbonyl compounds in good to excellent yields.

Sll0396 regulates transcription of the phycocyanin genes in Synechocystis sp. PCC 6803

  • Oh, In-Hye;Kim, Ho-San;Chung, Young-Ho;Kim, Young-Hye;Park, Young-Mok
    • Plant Biotechnology Reports
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    • 제4권3호
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    • pp.193-199
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    • 2010
  • An olive-green mutant was generated in Synechocystis sp. strain PCC 6803 by inactivation of the sll0396 gene. Whole-cell absorption spectra of the mutant revealed the missing of phycocyanin peak. An investigation of the low-temperature fluorescence emission spectra revealed that the $sll0396{\Omega}$ mutant has a reduced amount of phycocyanin. Western blot analysis showed that the mutant contained less phycocyanin ${\beta}$- and ${\alpha}$-subunits and lacked the 30- and 32-kDa linker polypeptides, and northern blot analysis revealed that the transcription of the 1.4-kb cpcBA gene encoding the phycocyanin ${\beta}$- and ${\alpha}$-subunits was lower in the mutant. The Sll0396 protein has a DNA-binding motif and shares homology with known response regulators. Our results indicate that Sll0396 plays a regulatory role in the transcription of the phycocyanin genes during phycobilisome synthesis.

Identification of Mutations in Protein Kinase CKIIβ Subunit That Affect Its Binding to Ribosomal Protein L41 and Homodimerization

  • Ahn, Bong-Hyun;Lee, Ji-Hoon;Bae, Young-Seuk
    • BMB Reports
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    • 제36권4호
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    • pp.344-348
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    • 2003
  • Protein kinase CKII is composed of two catalytic ($\alpha$ or $\alpha$') subunits and two regulatory ($\beta$) subunits. The $CKII{\beta}$ subunit is thought to mediate the tetramer formation and interact with other target proteins. However, its physiological function remains obscure. In this study, point mutants of $CKII{\beta}$ that are defective for the L41 binding were isolated by using the reverse two-hybrid system. A sequence analysis of the point mutants revealed that Asp-26, Met-52, and Met-78 of $CKII{\beta}$ are critical for L41 binding; Asn-67 (and/or Lys-139) and Met-52 are important for $CKII{\beta}$ homodimerization. Two point mutants, R75 and R83, of $CKII{\beta}$ interacted with L5, topoisomerase $II{\beta}$, and CKBBP1/SAG, but not with the wild-type $CKII{\beta}$. This indicates that $CKII{\beta}$ homodimerization is not a prerequisite for its binding to target proteins. These $CKII{\beta}$ point mutants may be useful in exploring the biochemical physiological functions of $CKII{\beta}$.

Curcumin suppresses the production of interleukin-6 in Prevotella intermedia lipopolysaccharide-activated RAW 264.7 cells

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • 제41권3호
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    • pp.157-163
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    • 2011
  • Purpose: Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. Methods: LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. $I{\kappa}B-{\alpha}$ degradation, nuclear translocation of NF-${\kappa}B$ subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-${\kappa}B$ was also analyzed. Results: Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of $I{\kappa}B-{\alpha}$ induced by P. intermedia LPS. Curcumin blocked NF-${\kappa}B$ signaling through the inhibition of nuclear translocation of NF-${\kappa}B$ p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. Conclusions: Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

베트남 지방수집종 벼의 종실특성변이 (Variation of Grain Traits in Landraces of Rice Collected from Vietnam)

  • 김현호;조재성
    • 농업과학연구
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    • 제24권2호
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    • pp.121-125
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    • 1997
  • 베트남으로부터 수집된 총 437 벼 지역종을 사용하여 총 종자 단백질을 분석하기 위하여 SDS-PAGE와 phenol 반응을 수행하였다. 세 가지 다른 형태의 glutelin a subunit이 검출되었다. 60kDa의 분자량인 wx 단백질 수준은 세 그룹으로 분리되었는데, 이는 non-glutelin, intermediate, 그리고 glutelin 전분 유형이다 종자 저장 단백질과 wx 단백질의 변이에 의한 실험결과 지역종은 7그룹으로 구분된다. Glutelin a subunit의 A와 B형의 빈도분포는 벼 지역종이 수집된 위도에 따라서 변하였다. 페놀 반응에 대한 지리학적인 구배가 검출되었다.

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Cloning of pdh Genes Encoding Subunits of Pyruvate Dehydrogenase Complex from Lactobacillus reuteri ATCC 55739

  • Nam, Su-Jin;Park, Jae-Yong;Kim, Jung-Kon;Ha, Yeong-Lae;Yun, Han-Dae;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제14권1호
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    • pp.197-201
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    • 2004
  • A 2-D gel protein analysis of Lactobacillus reuteri ATCC 55739 produced spots corresponding to subunits of the pyruvate dehydrogenase complex, as identified by N-terminal protein sequencing. Oligonucleotide probes specific for the subunits of the pyruvate dehydrogenase complex were synthesized ,md used to screen a L. reuteri genomic library to clone the structural genes. Two positive clones were isolated and identified as having the same 2.2 kb insert. A pdhB encoding the $\beta$-subunit of El subunit (pyruvate dehydrogenase component) of the pyruvate dehydrogenase complex was located in the middle of the insert. Furthermore, a 5' truncated pdhA encoding the $\alpha$-subunit of the E1 subunit and a 3' truncated pdhC encoding the E2 subunit (dihydrolipoamide acetyltransferase) were also located upstream and downstream of the pdhB, respectively.

Expressions of Gonadotropin Subunit Genes in the Reproductively Inactive Golden Hamsters

  • Choi, Donchan
    • 한국발생생물학회지:발생과생식
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    • 제26권2호
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    • pp.37-47
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    • 2022
  • Photoperiod has well been established to regulate testicular activities in golden hamsters. These animals breed actively around summer but become infertile in winter. In males, testicles are full of multistep germ cells including spermatozoa in summer. But in winter only fundamental cells consisting of the testicles are detected. The testicular degeneration is accompanied by the reduced levels of blood gonadotropins and testosterone. In this study, the expressions of gonadotropin subunit genes were investigated in the reproductive active and inactive testicles. And parts of sequences of the gonadotropin subunits were identified and compared with those of other rodents. As results, common gonadotropin alpha (CGa), follicle-stimulating hormone (FSH) β, and luteinizing hormone (LH) β genes were equivalently detected in pituitaries of both sexually active and inactive animals. In considering low concentrations of gonadotropin hormones determined in pituitary, the present findings imply that the processes involved in translation and/or formation of functional hormones could be impeded in the sexually inactive hamsters. All the nucleotide sequences of gonadotropin subunits identified in this study were same as those reported previously except for one base in CGa. An unsure amino acid deduced from the CGa sequence was confirmed from mRNA sequencing. The outcomes mentioned above suggest that animals with regressed testes prepare for the sexually active period forthcoming in the future.

콩 돌연변이 계통의 단백질 특성 (Seed Protein Quality of Soybean Mutants)

  • 양무희
    • 한국작물학회지
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    • 제39권3호
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    • pp.278-284
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    • 1994
  • 콩단백질의 황 아미노산함량은 가축 영양학상 중요한 위치를 차지하기 때문에 신계통이 가져야만 할 필수조건일지도 모른다. 콩 계통간에 저장단백질의 유전적변이가 존재한다면 이는 기존의 육종방법을 통하여 콩의 종자단백질 구성성분을 유전적으로 변경하여 품질을 개량할 수 있는 가능성을 시사하고 있다. 본 연구는 여러 문헌에 보고된 콩종자 저장단백질의 돌연변이 계통들을 선별하여 콩단백질의 품질을 향상시키기 위한 육종 재료로서의 가능성을 평가하기 위하여 실행되었다. 수집된 돌연변이 계통들은 저장단백질의 또 다른 특성을 나타내었다. 그 돌연변이 계통들 중에서 Keburi(P.I.417016), Keburi(P.I.506817), P.I.154608-1 등은 황 아미노산 함량이 상대적으로 다른 돌연변이 계통보다 높은 1.9, 2.1, 1.8%를 나타내었으며, 이는 7S 단백질인 ${\alpha}$ ', ${\alpha}$ , ${\beta}$단백질 함량이 상대적으로 낮기 때문인 것으로 나타났다. 그러므로 그 돌연변이 계통들 중에서 Keburi(P.I.417016), Keburi(P.I.506817), P.I.54608-1 등은 황 아미노산 함량을 향상시키기 위한 중요한 육종재료로, 그 외 돌연변이 계통들은 다른 용도의 육종 재료로 이용할 수 있을 것으로 추측된다.

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Biosynthesis of a Biological Active Single Chain Equine Chorionic Gonado-tropin

  • Min, Kwan-Sik
    • Journal of Life Science
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    • 제11권2호
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    • pp.103-107
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    • 2001
  • The equine chorionic gonadotropin (eCG) subunits $\alpha$ and ${\beta}$ are transcribed from different genes and associate noncovalently to form the bioactive eCG heterodimer. Dimerization is rate limiting for eCG secretion, and dissociation leads to hormone inactivation. The correct conformation of the heterodimer is alto important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. To determine whether ${\alpha}$ and ${\beta}$ subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule by fusing the carboxyl terminus of the eCG ${\beta}$-subunit to the amino terminus of the af-subunit was construe-ted and transfected into chinese hamster ovary (CHO-Kl) cells. LH- and FSH-like activities were assayed in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells and granulosa cells, respectively. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG ${\alpha}$/${\beta}$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG ${\alpha}$/${\beta}$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-K1 cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models of FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

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Synthetic Lead Compounds Modulate Activity of Large-conductance $Ca^{2+}$-activated Potassium Channels Expressed in Xenopus Oocytes

  • Ha, Tal-Soo;Kim, Yong-Chul;Park, Chul-Seung
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.42-42
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    • 2003
  • Large-conductance $Ca^{2+}$-activated potassium channels ($BK_{Ca}$ are a widely distributed and play key roles in various cell functions. In nerve cells, $BK_{Ca}$ channels shorten the duration of action potentials and block $Ca^{2+}$ entry thereby repolarizing excitable cells after excitation. $BK_{Ca}$ channel opening has been postulated to confer neuroprotection during stroke, and has attracted attention as a means for therapeutic intervention in asthma, hypertension, convulsions, and traumatic brain injury. Several natural and synthetic compounds including a steroid hormone, $\beta$-estradiol, have been identified as the activators of $BK_{Ca}$ channels. Based on the structural features of the previously reported activators of $BK_{Ca}$ channels, we designed several lead compounds, synthesized chemically, and tested their functional activity on cloned $BK_{Ca}$ channels. The $\alpha$ subunit of rat $BK_{Ca}$ channel was expressed alone or with different $\beta$ subunits in Xenopus oocytes and the effects of the compounds were tested electrophysiological means. One of the lead compounds affected the activity of the $\alpha$ subunit of $BK_{Ca}$ channel in a $\beta$ subunit-specific manner. While the activity of B $K_{ca}$ channel $\alpha$ subunit was Potentiated, the channel composed of $\alpha$ and $\beta$1 subunits were inhibited by this compound. We are currently investigating the mechanism of the $\beta$ subunit-dependent effects and planning to localize the receptor site of the lead compound.f the lead compound.

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