• 제목/요약/키워드: Site-specificity

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Substitution of Asp-223 Residue to Leu in Yeast Alcohol Dehydrogenase and Coenzyme Specificity (효모 알코올 탈수소효소 아스파르트산-223 잔기의 루신으로 치환과 보조효소의 특이성)

  • Lee, Kang-Man;Ryu, Ji-Won
    • YAKHAK HOEJI
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    • v.36 no.5
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    • pp.469-473
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    • 1992
  • Yeast alcohol dehydrogenase (YADH) has an acidic residue that interacts with the 2'- and 3'-hydroxyl groups of the adenosine ribose of the $NAD^+$ coenzyme. The acidic residue of Asp-223 (according to horse liver alcohol dehydrogenase amino acid sequence) is supposed to determine the coenzyme specificity for $NAD^+$ rather than $NADP^+$. We mutated Asp-223 to leucine and the mutant YADH was expressed in yeast and characterized for the coenzyme specificity. The turnover numbers of mutant enzyme for $NAD^+$ and ethanol were decreased 3.5- and 4.8-fold compared to wild-type enzyme, respectively. Contrastively, catalytic specificity for $NADP^+$ was increased 13-fold. As a result, the mutant YADH also employed $NADP^+$ as a coenzyme.

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Improved Detection of Helicobacter pylori Infection and Premalignant Gastric Mucosa Using "Site Specific Biopsy": a Randomized Control Clinical Trial

  • Tongtawee, Taweesak;Dechsukhum, Chavaboon;Leeanansaksiri, Wilairat;Kaewpitoon, Soraya;Kaewpitoon, Natthawut;Loyd, Ryan A;Matrakool, Likit;Panpimanmas, Sukij
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8487-8490
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    • 2016
  • Background: Helicobacter pylori infection and premalignant gastric mucosa can be reliably identified using conventional narrow band imaging (C-NBI) gastroscopy. The aim of our study was to compare standard biopsy with site specific biopsy for diagnosis of H. pylori infection and premalignant gastric mucosa in daily clinical practice. Materials and Methods: Of a total of 500 patients who underwent gastroscopy for investigation of dyspeptic symptoms, 250 patients underwent site specific biopsy using C-NBI (Group 1) and 250 standard biopsy (Group 2). Sensitivity, specificity, and positive and negative predictive values were assessed. The efficacy of detecting H. pylori associated gastritis and premalignant gastric mucosa according to the updated Sydney classification was also compared. Results: In group 1 the sensitivity, specificity, positive and negative predictive values for predicting H. pylori positivity were 95.4%, 97.3%, 98.8% and 90.0% respectively, compared to 92.9%, 88.6%, 83.2% and 76.1% in group 2. Site specific biopsy was more effective than standard biopsy in terms of both H. pylori infection status and premalignant gastric mucosa detection (P<0.01). Conclusions: Site specific biopsy using C-NBI can improve detection of H. pylori infection and premalignant gastric mucosa in daily clinical practice.

Substrate specificity of bacterial endoribonuclease toxins

  • Han, Yoontak;Lee, Eun-Jin
    • BMB Reports
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    • v.53 no.12
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    • pp.611-621
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    • 2020
  • Bacterial endoribonuclease toxins belong to a protein family that inhibits bacterial growth by degrading mRNA or rRNA sequences. The toxin genes are organized in pairs with its cognate antitoxins in the chromosome and thus the activities of the toxins are antagonized by antitoxin proteins or RNAs during active translation. In response to a variety of cellular stresses, the endoribonuclease toxins appear to be released from antitoxin molecules via proteolytic cleavage of antitoxin proteins or preferential degradation of antitoxin RNAs and cleave a diverse range of mRNA or rRNA sequences in a sequence-specific or codon-specific manner, resulting in various biological phenomena such as antibiotic tolerance and persister cell formation. Given that substrate specificity of each endoribonuclease toxin is determined by its structure and the composition of active site residues, we summarize the biology, structure, and substrate specificity of the updated bacterial endoribonuclease toxins.

Identification of Substrate Specificity Determinant of Achromobacter Protease I (API) and Catalytic Activity of Mutant D225E for Ornithine Substrate (Achromobacter Protease I (API)의 기질특이성 결정기의 동정과 변이체[D225E]의 Ornithine 기질에 대한 촉매활성)

  • Lim, Seong-Il;Kwon, Oh-Jin;Choi, Cheong
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.189-195
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    • 1997
  • The structural basis of Iysine specificity of Achromobacter protease I (API) was investigated by means of site-directed mutagenesis. The precursor protein in which Glu190, one of the two candidates for determining Iysine specificity, was substituted by glutamine, aspartic acid or leucine was processed autocatalytically to attaln full pretense activity with lysine specificity. The substitution of the other candidate, Asp225, for asparagine or leucine produced no mature active forms of pro-API. The precursor protein of the mutant D225E slowly matured autocatalytically. The lysylendopeptidase activity of the mature D225E was 0.25% of that of native API, and this reduced activity is mainly due to a decrease in the affinity of the enzyme for lysine. These results suggest that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase. However, D225E exhibited no measurable activity for synthetic ornithine substrate. Since the hydroxyl group of Ser194 in this mutant retained essentially the same reactivity to DFP or PMSF as that in native API, it can be noted that a methylene unit longer side chain of residue 225 is not compensated by a methylene unit shorter side chain at subsite P1 in the bound substrate.

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Accuracy of Preoperative Urinary Symptoms, Urinalysis, Computed Tomography and Cystoscopic Findings for the Diagnosis of Urinary Bladder Invasion in Patients with Colorectal Cancer

  • Woranisarakul, Varat;Ramart, Patkawat;Phinthusophon, Kittipong;Chotikawanich, Ekkarin;Prapasrivorakul, Siriluck;Lohsiriwat, Varut
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7241-7244
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    • 2014
  • Background: To determine the accuracy of preoperative urinary symptoms, urinalysis, computed tomography (CT) and cystoscopic findings for the diagnosis of urinary bladder invasion in patients with colorectal cancer. Materials and Methods: Records of patients with colorectal cancer and a suspicion of bladder invasion, who underwent tumor resection with partial or total cystectomy between 2002 and 2013 at the Faculty of Medicine Siriraj Hospital, were reviewed. Correlations between preoperative urinary symptoms, urinalysis, cystoscopic finding, CT imaging and final pathological reports were analyzed. Results: This study included 90 eligible cases (71% male). The most common site of primary colorectal cancer was the sigmoid colon (44%), followed by the rectum (33%). Final pathological reports showed definite bladder invasion in 53 cases (59%). Significant features for predicting definite tumor invasion were gross hematuria (OR 13.6, sensitivity 39%, specificity 73%), and visible tumor during cystoscopy (OR 5.33, sensitivity 50%, specificity 84%). Predictive signs in CT imaging were gross tumor invasion (OR 7.07, sensitivity 89%, specificity 46%), abnormal enhancing mass at bladder wall (OR 4.09, sensitivity 68%, specificity 66%), irregular bladder mucosa (OR 3.53, sensitivity 70%, specificity 60% ), and loss of perivesical fat plane (OR 3.17, sensitivity 81%, specificity 43%). However, urinary analysis and other urinary tract symptoms were poor predictors of bladder involvement. Conclusions: The present study demonstrated that the most relevant preoperative predictors of definite bladder invasion in patients with colorectal cancer are gross hematuria, a visible tumor during cystoscopy, and abnormal CT findings.

A Study on the Place Specificity of Graffiti Arts Used as Fusion Design Elements in Urban Regeneration (융합 디자인 요소로서 도시재생에 활용된 그라피티의 장소특정성에 관한 연구)

  • Lim, Byung-Woo
    • Journal of Digital Convergence
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    • v.15 no.11
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    • pp.455-461
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    • 2017
  • This study aims to clarify how the graffiti arts used in urban regeneration projects have come to possess the place specificity as public arts. For the research, the theoretical background of the place specificity of graffiti art was explored with the understanding of the concept of place specificity of public art, before comparing the difference of graffiti arts as public art applied to the urban regeneration projects, depending on their installation location and artist. In addition, the specificity of the place where graffiti artwork was installed and its significance in the context of urban regeneration project were explored. The case study is limited to graffiti arts installed in Bosan-dong, Dongducheon-si, where several urban regeneration projects have been pushed for since 2015. It was found from the analysis that the place specificity of individual art works is maximized when they are merged with cultural contents reflecting the history and place specificity of target cities, such as design, crafts, music and festivals, rather than with the place specificity of the works of individual artists. The result of this study suggests that when graffiti arts are fused with urban cultural contents as fusion design elements, their place specificity is maximized enough to elevate them to the level of public arts, and enable them to function as the media of urban regeneration.

Analysis of Active Center in Hyperthermophilic Cellulase from Pyrococcus horikoshii

  • Kang, Hee-Jin;Ishikawa, Kazuhiko
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1249-1253
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    • 2007
  • A hyperthermostable endoglucanase from Pyrococcus horikoshii with the capability of hydrolyzing crystalline cellulose was analyzed. A protein engineering study was carried out to obtain a reduced-size mutant. Five amino acid residues at both the N- and C-terminus were found to be removable without any loss of activity or thermal stability. Site-directed mutagenesis was also performed on R102, N200, E201, H297, Y299, E342, and W377, residues possibly involved in the active center or in the recognition and binding of a cellulose substrate. The activity of the resulting mutants was considerably decreased, confirming that the mutated residues were all important for activity. A reduced-size enzyme, as active as the wild-type endoglucanase, was successfully obtained, plus the residues critical for its activity and specificity were confirmed. Consequently, an engineered enzyme with a reduced size was obtained, and the amino acids essential for activity were confirmed by site-directed mutagenesis and comparison with a known three-dimensional structure.

The Features of the Pavilions, Follies, and Installations of the Glass House (글라스하우스의 파빌리온, 폴리, 인스톨레이션의 특성)

  • Kim, Ran-Soo
    • Journal of architectural history
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    • v.26 no.1
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    • pp.71-82
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    • 2017
  • Pavilions, follies, and installations provide a place with a new experience beyond that of simple garden architecture. From this point of view, this study tried to analyze the constructs in the Glass House site, which Philip Johnson has built for 50 years. After Chapter 1 Introduction, which summarized the background of the study, Chapter 2 investigated the design background of the landscape and the types of the constructs there. It also, studying literature on pavilions, follies, and installations, defined the basic meanings of them. Chapter 3 identified the features of each construct through the case studies of it, analyzing Johnson's intentions on it. These features are such as the applications of classical follies, the quotations of architectural history, fusion with art, architectural experiments, and the monuments of personal history. In conclusion, this study, finding the site specificity as a common feature of pavilions, follies, and installations, referred to two aspects of this, which are not only physical placeness but also cultural media.

A Method for Identifying Splice Sites and Translation Start Sites in Human Genomic Sequences

  • Kim, Ki-Bong;Park, Kie-Jung;Kong, Eun-Bae
    • BMB Reports
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    • v.35 no.5
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    • pp.513-517
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    • 2002
  • We describe a new method for identifying the sequences that signal the start of translation, and the boundaries between exons and introns (donor and acceptor sites) in human mRNA. According to the mandatory keyword, ORGANISM, and feature key, CDS, a large set of standard data for each signal site was extracted from the ASCII flat file, gbpri.seq, in the GenBank release 108.0. This was used to generate the scoring matrices, which summarize the sequence information for each signal site. The scoring matrices take into account the independent nucleotide frequencies between adjacent bases in each position within the signal site regions, and the relative weight on each nucleotide in proportion to their probabilities in the known signal sites. Using a scoring scheme that is based on the nucleotide scoring matrices, the method has great sensitivity and specificity when used to locate signals in uncharacterized human genomic DNA. These matrices are especially effective at distinguishing true and false sites.

Binding Subsites In the Active Site of $Zn^{2+}$-Glycerophosphocholine Cholinephosphodiesterase

  • Sok, Dai-Eun;Kim, Mee-Ree
    • BMB Reports
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    • v.28 no.2
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    • pp.94-99
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    • 1995
  • The properties of binding sites in the active site of $Zn^{2+}$-glycerophosphocholine cholinephosphodiesterase were examined using substrates and inhibitors of the enzyme. Phosphodiesterase hydrolyzed p-nitrophenylphosphocholine, p-aminophenylphosphocholine, and glycerophosphocholine, but did not hydrolyze either acylated glycerophosphocholine or bis (p-nitrophenyl)phosphate, suggesting a size limitation for interaction with a glyceryl moiety-binding subsite. The hydrolysis of p-nitrophenylphosphocholine was competitively inhibited by glycerophosphocholine and p-aminophenylphosphocholine, while glycerophosphoethanolamine was a weak inhibitor. The enzyme was also inhibited by choline, but not by ethanolamine. Thiocholine, a much more potent inhibitor than choline, was more inhibitory than cysteamine, suggesting a strict specificity of an anionic subsite adjacent to a $Zn^{2+}$ subsite. Of all oxyanions tested, the tellurite ion was found to strongly inhibit the enzyme by binding to a $Zn^{2+}$ subsite. The inhibitory role of tellurite was synergistically enhanced by tetraalkylammonium salts, but not by glycerol. Deactivation of the enzyme by diethylpyrocarbonate was partially protected by choline, but not by glycerophosphate. It is suggested that the active site of phosphodiesterase contains three binding subsites.

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