• Title/Summary/Keyword: SH2

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Sclerotiorin and Isochromophilone IV: Inhibitors of Grb2-Shc Interaction, Isolated from Penicillium multicolor F1753

  • Nam, Ji-Youn;Son, Kwang-Hee;Kim, Hyae-Kyeong;Han, Mi-Young;Kim, Sung-Uk;Choi, Jung-Do;Kwon, Byoung-Mog
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.544-546
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    • 2000
  • Grb2 is an important adaptor protein in the mitogenic Ras signaling pathway of receptor tyrosine kinases, and contains one SH2 domain and two SH3 domains. The SH2 domain binds to specific phosphotyrosine motifs on receptors or adaptor proteins such as Shc. The SH2 domain antagonists may lead to blocking of the oncogenic Ras signals and to developing new antitumor agents. In the course of screening SH2 antagonists from natural sources, cslerotiorin (1) and isochromophilone IV (2) were isolated from a strain, Penicillium multicolor F1753, and their structures were established by NMR spectral data. The metabolites significantly inhibited the binding between the Grb2-SH2 domain and phosphopeptide derived from the Shc protein, with $IC_{50}$ values of $22{\;}\mu\textrm{M}{\;}and{\;}48{\;}\mu\textrm{M}$ for (1) and (2), respectively. The compounds are the first nonpeptidic inhibitors of the SH2 domain from a natural source.

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Expression, Purification and NMR studies of SH3YL1 SH3 domain

  • Shrestha, Pravesh;Yun, Ji-Hye;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
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    • v.14 no.2
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    • pp.105-116
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    • 2010
  • SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of $\beta$-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.

Effects of Bupleuri radix Extract on Axon Regrowth in the Injured Sciatic Nerve of Rats (흰쥐의 좌골신경축삭 압좌 손상 후 시호(柴胡) 추출물에 의한 재생반응성 개선효과)

  • Kang, Jun-Hyuk;Oh, Min-Seok
    • The Journal of Korean Medicine
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    • v.31 no.1
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    • pp.93-111
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    • 2010
  • Objectives: The present study was performed to evaluate the potential effects of Bupleuri radix (SH) on regenerative activities in the peripheral sciatic nerve after crushing injury in rats. Methods: Axonal regeneration after crush injury in rats was analyzed by immunofluorescence staining using anti-NF-200 antibody and retrograde tracing of DiI-axons. Changes in protein levels in the sciatic nerve axons and DRG tissue were analyzed by Western blot analysis and immunofluorescence staining. Effects of SH extract treatment on neurite outgrowth was examined by immunofluorescence staining for cultured DRG neurons. Results: Major findings on the effects of SH extract treatment on axonal regeneration are summarized as follows. 1. SH-mediated enhancement in axonal regeneration was identified by immuno- fluorescence straining of NF-200 protein and retrograde tracing of DiI-labeled axons. 2. Axonal GAP-43 protein levels were upregulated by SH not only in the injured axons but also in the DRG sensory neurons corresponding to sciatic sensory axons. 3. Phospho-Erk1/2 protein levels were increased in both injured axonal area and DRG sensory neurons by SH. Phospho-Erk1/2 was also found in non-neuronal cells in the injured axons. 4. SH elevated levels of Cdc2 protein produced in Schwann cells in the distal portions of injured sciatic nerves. 5. The neurite outgrowth of DRG sensory neurons in culture was augmented by SH, and these changes were positively associated with GAP-43 production levels in the DRG neurons. Conclusions: These data suggest that SH extract improves the regenerative responses of injured peripheral neurons, and thus may be useful for understanding molecular basis for the development of therapeutic strategies.

MeSH Semi Indexing of the Korean Biomedical Literature, using NLM Medical Text Indexer (NLM Medical Text Indexer를 활용한 우리나라 의학문헌의 MeSH Semi Indexing 방안)

  • Jeong, Sona;Lee, Choon Shil
    • Proceedings of the Korean Society for Information Management Conference
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    • 2010.08a
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    • pp.21-28
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    • 2010
  • 본 연구에서는 PubMed에 등재되었으나 Medical Subject Headings(MeSH)가 부여되지 않은 국내 의학학술지의 문헌을 대상으로 미국국립의학도서관 (NLM: National Library of Medicine)의 Medical Text Indexer(MTI)를 활용하여 MeSH 용어를 추천받은 후, PubMed 레코드의 유사주제문헌 (Relation Citations, PRC)에 부여된 MeSH와의 일치여부를 분석하였다. 또한 논문의 저자가 부여한 키워드(저자키워드)와 PRC MeSH의 일치여부도 비교하였다. PRC MeSH와 MTI MeSH 추천어의 일치율은 주표목이 21.1%였고, 체크태그는 18.1%, 부표목은 16.5%로 나타났다. 우리나라 의학논문에 나타난 저자키워드의 중요한 특징은 MeSH 주표목 위주이고, 체크태그와 부표목은 거의 사용하지 않는 것이다. 따라서 저자키워드와 PRC MeSH 주표목과의 일치율은 23.4%에 이르지만, 체크태그와 부표목의 일치율은 각각 1%, 2.1%였다. 색인전문가가 통제어휘를 사용하여 색인하는 과정에서 PRC와 MTI의 MeSH 주표목과 저자키워드가 일치하는 용어를 주표목으로 부여하고, PRC와 MTI가 추천하는 체크태그와 부표목을 활용하는 등 국내 의학문헌의 MeSH 용어 부여 작업을 반자동화(semi-indexing)하면, 정확하고 신속한 MeSH 부여 작업이 가능할 것이다.

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Effect of pH on the Production and Characteristics of Protease by Bacillus sp. SH-8 and Bacillus sp. SH-8M (Bacillus sp. SH-8과 Bacillus SP. SH-8M의 Protease 생산 및 특성에 미치는 pH의 영향)

  • Shim, Chang-Whan;Jeong, Kwang-Seon;Shin, Won-Cheol;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.22 no.1
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    • pp.59-64
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    • 1994
  • The production and the characteristics of protease produced by Bacillus sp. SH-8 and Bacillus sp. SH-8M were investigated under the different pH conditions. Bacillus sp. SH-8 and Bacillus sp. SH-8M showed the maximum activity of protease at 60 hours(70 units/ml) AND 96 houre(50 units/ml) cultivation, respectively, under the alkaline condition(pH 10.2). However, Bacillus sp. SH-8M exhibited the maximum activities in 8 days cultivation at pH 6.9 and in 6 days cultivation at pH 7.7 Bacillus sp. SH-8M showed the protease activity at the pH change from alkaline to neutral condition, whereas Bacillus sp. SH-8 did not. In addition. all the enzymatic characteristics of protease produced by Bacillus sp. SH-8 and Bacillus sp. SH-8M were similar with the regardless of different pH conditions.

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Cultural Characterization of Bacteriolytic Bacillus subtilis SH-1 Isolated from Pusan Coastal Sea (해양에서 분리한 용균세균인 Bacillus subtilis SH-1의 배양특성)

  • 류병호;진성현
    • Journal of Food Hygiene and Safety
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    • v.10 no.4
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    • pp.231-237
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    • 1995
  • Bacillus subtilis SH-1 have been isolated and identified from coastal sea, in Pusan, The optimal cultural characterization of Bacillus subtilis SH-1 for 속 production of bacteriolytic enzyme was determained. Bacillus subtilis SH-1 produced the bacteriolytic enzyme well in the medium consist of 1.0% glucose, 1.0% yeast extract, 1.0% NaCI, 0.02% $K_2HPO_4,\;0.002%\;MgSo_4{\cdot}7H_2O,\;0.001%\;MnSO_4{\cdot}5H_2O,\;0.0001%\;FeSO_4{\cdot}7H_2O$. The optimal medium pH, incubation temperature, and shaking tome for the highest production of the enzyme were 8.0, $30^{\circ}C$ and 28 hours respectively.

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Molecular interaction between SH3 domain of PACSIN2 and proline-rich motifs of Cobll1

  • Yoo, Hee-Seop;Seok, Seung-Hyeon;Kim, Ha-Neul;Kim, Ji-Hun;Seo, Min-Duk
    • Journal of the Korean Magnetic Resonance Society
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    • v.26 no.3
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    • pp.34-39
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    • 2022
  • The SH3 domain found within a variety of proteins is comprised of generally 60 residues, and participated in protein-protein interactions with proline-rich motifs. Cobll1 was identified as a distinct molecular marker associated with CML progression, and PACSIN2 was discovered a novel Cobll1 binding partner through direct interaction between a SH3 domain of PACSIN2 and three proline-rich motifs of Cobll1. To understand the structural basis of interactions between PACSIN2 and Cobll1, backbone assignments of PACSIN2 SH3 domain were performed. Furthermore, three proline-rich peptides of Cobll1 were titrated to 15N-labeled PACSIN2 SH3 domain in various ratios. Our chemical shift changes data and conserved SH3 sequence alignment will be helpful to analyze fundamental molecular basis related to the interaction between PACSIN2 and Cobll1.

A Screening Method for Src Homology 3 Domain Binding Blockers Based on Ras Signaling Pathway

  • Ko, Woo-Suk;Yoon, Sun-Young;Kim, Jae-Won;Lee, Choong-Eun;Han, Mi-Young
    • BMB Reports
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    • v.30 no.5
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    • pp.303-307
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    • 1997
  • Grb2, which is composed of a Src homology 2 (SH2) domain and two Src homology 3 (SH3) domains, is known to serve as an adaptor protein in signaling for Ras activation. Thus, a blocker of the Grb2 interactions with other proteins can be a potential candidate for an anticancer drug. In this study, we have developed a high throughput screening method for SH3 domain binding ligands and blockers. Firstly, we made and purified the glutathione S-transferase (GST)-fusion proteins with the Grb2 SH2 and SH3 domains, and the entire Grb2. This method measures the binding of a biotin-labeled oligopeptide, derived from a Grb2/SH3 binding motif in the hSos, to the GST-fusion proteins, which are precoated as glutathione S-transferase fusion protein on a solid phase. When $1\;{\mu}g$ of each fusion protein was used to coat the wells, both N- and C- terminal SH3 the domains as well as the whole of Grb2 were able to interact with the biotin-conjugated ligand peptide, while the SH2 domain and GST alone showed no binding affinity. Although N- and C- terminal SH3 domains showed an increase of binding to the ligand peptide in proportion to the amount of peptide, the GST fusion protein with Grb2 demonstrated much higher binding affinity. GST-Grb2 coating on the solid phase showed a saturation curve; 66 and 84% of the maximal binding was observed at 100 and 300 ng/$100\;{\mu}l$, respectively. This binding assay system was peptide sequence-specific, showing a dose-dependent inhibition with the unlabeled peptide of SH3 binding motif. Several other peptides, such as SH2 domain binding motifs and PTB domain binding motif, were ineffective to inhibit the binding to the biotin-conjugated ligand peptide. These results suggest that our method may be useful to screen for new anticancer drug candidates which can block the signaling pathways mediated by SH3 domain binding.

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Studies on the Isolation and Utilization of Apple Wine Yeasts (우수 사과주효모(酒酵母)의 분리(分離)와 이용(利用)에 관(關)한 연구(硏究))

  • Park, Yoon-Joong;Kim, Chan-Jo;Lee, Suk-Kun;Oh, Man-Jin;Sohn, Cheon-Bae
    • Korean Journal of Agricultural Science
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    • v.5 no.1
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    • pp.35-41
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    • 1978
  • Extensive selection works on wild yeasts of fruits were carried cut to obtain strains which are applicable to apple wine making. Among the total number of 1,358 yeast strains which were isolated from various fruit samples collected from the vicinity of Daejeon and other regions cf Korea, the strains SH-49, SH-129 and SH-338 were found to be useful. Then experiments on their morphological and physiological characteristics, and on the aspects of practical use in apple wine making were proceeded. The results obtained were as follows: 1. The strains SH-49 and SH-129, particularly SH-49, were appeared to have good fermentation ability, tolerance to sulfur dioxide and to produce fine quality of apple wine. 2. Apple wines made by using the strain, SH-49 and SH-129 contained less amount of total acids than those by other strains. 3. Apple wines of SH-49 and SH-129 were clarified rapidly during the primary fermentation period, and their absorbancy at 430 nm after 45 days of storage were approximately half of others. 4. Apple wine of SH-338 contained higher amount of residual sugar and its quality was superior to others. It is considered that this strain could be used in the production of apple wine of a characteristic quality. 5. The strains SH-49 and SH-338 were identified as a Saccharomyces cerevisiae according to Taxanomic Study of Yeasts by Lodder, however, classification of SH-129 was suspended for further study.

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Evaluation of Dietary Supplementation of Sargassum horneri for Pacific White Shrimp Litopenaeus vannamei (흰다리새우(Litopenaeus vannamei) 사료 내 괭생이모자반(Sargassum horneri) 분말의 이용가능성 평가)

  • Eom, Gunho;Shin, Jaebeom;Lim, Se-Jin;Lee, Kyu-Tae;Han, Young-Seok;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.6
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    • pp.909-917
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    • 2020
  • This study was conducted to evaluate dietary supplementation of Sargassum horneri (SH) meal on growth performance, feed utilization, innate immunity, antioxidant capacity and disease resistance of Pacific white shrimp. The diets were added with graded levels of SH meal by 0, 0.5, 1, 2, 4 and 8% (designated as Con, SH0.5, SH1, SH2, SH4 and SH8, respectively). Quadruplicate groups of shrimp were hand-fed with one of the diets five times daily for 39 days. The innate immunity and antioxidant capacity of shrimp were significantly improved by the dietary SH supplementation. The cumulative survival of shrimp exposed to Vibrio parahaemolyticus in a challenge was higher in shrimp group fed SH diets, except for SH8, than that of shrimp fed the control diet. Growth performance and feed utilization of the shrimp were significantly decreased with the dietary SH inclusions except for 0.5%. Results indicated that dietary SH meal could be used as a functional supplement for improvements in innate immunity, antioxidant capacity and disease resistance in Pacific white shrimp. The suggested dietary inclusion level of dried SH is approximately 0.5% for the shrimp.