• Title/Summary/Keyword: $3_{10}$-helix

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Study on the structure of cAMP receptor protein(CRP) by temperature change (온도변화에 의한 cAMP 수용성 단백질(CRP)의 구조)

  • 주종호;구미자;강종백
    • Journal of Life Science
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    • v.10 no.3
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    • pp.279-285
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    • 2000
  • CRP (cyclic AMP receptor protein) regulate transcription of catabolite-sensitive genes in Escherichia coli. Wild-type and mutant CRP (S83G and S128A) proteins were used to measure the thermal stability and the temperature-dependent structural change by proteolytic digestion, UV spectrophotometer and CD spectrapolarimeter. The result indicated that wild-type CRP was more thermally stable than the mutant CRPs in the presence of cAMP. At a low temperature, wild-type CRP with cAMP was more sensitive to subtilisin than the mutant CRPs. At a high temperature, there was no difference of sensitivity to subtilisin among wild-type, S83G and S128A CRPs. CD spectra suggested that the secondary structure of CRP was destroyed partially at a high temperature.

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Structure-Activity Relationships of 9-mer Antimicrobial Peptide analogue of Protaetiamycine, 9Pbw2

  • Kim, Jin-Kyoung;Lee, Eun-Jung;Jung, Ki-Woong;Kim, Yang-Mee
    • Journal of the Korean Magnetic Resonance Society
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    • v.15 no.1
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    • pp.1-13
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    • 2011
  • 9Pbw2 is a 9-mer analog of protaetiamycine derived from the larvae of the beetle Protaetia brevitarsis. Previously, we designed four 9-mer peptide analogues to optimize the balance between the hydrophobicity and cationicity of the peptides and to increase bacterial cell selectivity. Among them, 9Pbw2 has high antibacterial activity without cytotoxicity. The results obtained in previous study suggest that the bactericidal action of 9Pbw2 may be attributed to the inhibition of the functions of intracellular components after penetration of the bacterial cell membrane. In order to understand structure-activity relationships, we determined the three-dimensional structure of 9Pbw2 in 200 mM DPC micelle by NMR spectroscopy. 9Pbw2 has one hydrophobic turn helix from $Trp^3$ to $Arg^8$ and positively charged residues at the N- and C-terminus. This result suggested that positively charged residues from position at the C-terminus in 9Pbw2 may be important for the primary binding to the negatively charged phospholipid head groups in bacterial cell membranes and hydrophobic residues in the middle portion face toward the acyl chains of the hydrophobic lipid in the bacterial cell membrane.

A replication study of genome-wide CNV association for hepatic biomarkers identifies nine genes associated with liver function

  • Kim, Hyo-Young;Byun, Mi-Jeong;Kim, Hee-Bal
    • BMB Reports
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    • v.44 no.9
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    • pp.578-583
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    • 2011
  • Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are biochemical markers used to test for liver diseases. Copy number variation (CNV) plays an important role in determining complex traits and is an emerging area in the study various diseases. We performed a genome-wide association study with liver function biomarkers AST and ALT in 407 unrelated Koreans. We assayed the genome-wide variations on an Affymetrix Genome-Wide 6.0 array, and CNVs were analyzed using HelixTree. Using single linear regression, 32 and 42 CNVs showed significance for AST and ALT, respectively (P value < 0.05). We compared CNV-based genes between the current study (KARE2; AST-140, ALT-172) and KARE1 (AST-1885, ALT-773) using NetBox. Results showed 9 genes (CIDEB, DFFA, PSMA3, PSMC5, PSMC6, PSMD12, PSMF1, SDC4, and SIAH1) were overlapped for AST, but no overlapped genes were found for ALT. Functional gene annotation analysis shown the proteasome pathway, Wnt signaling pathway, programmed cell death, and protein binding.

Metabolites of Doxylamine succinate in Human Urine (인체 뇨중의 숙신산 독실아민의 대사체)

  • Eom, Khee-Dong;Jung, Byung-Hwa;Chung, Bong-Chul;Slikker, William;Park, Jong-Sei
    • YAKHAK HOEJI
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    • v.36 no.3
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    • pp.230-240
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    • 1992
  • The metabolic profile of doxylamine, N,N-dimethyl-2-[1-phenyl-1-(2-pyridinyl)ethoxy] ethanamine, was determined in the human urine. The free fractions of extracts were obtained without hydrolysis, and the conjugated fractions of extracts were obtained with enzyme hydrolysis using ${\beta}-glucuronidase/arylsulfatase$ from Helix pomatia. The mixture of acetic anhydride/pyridine (10 : 1, v : v) was used to derivatize the urinary extracts and then analyzed by gas chromatography and mass selective detector. N-desmethyldoxylamine, doxylamine carboxylic acid, desaminohydroxydoxylamine, N, N-didesmethyldoxylamine, N-acetyl conjugates of N-desmethyl and N, N-didesmethyldoxylamine, quarternary ammonium N-glucuronide of doxylamine, N-desmethyldoxylamine N-glucuronide and unchanged doxylamine were detected in the human urine obtained after oral treatment with doxylamine succinate. $N-methyl-{\alpha}-hydroxy-2-[1-phenyl-1-(2-pyridinyl)$ ethoxy] ethanamine, which can be a key intermediate of this metabolism, was tentatively identified by the interpretation of its mass spectrum. In this study, we proposed the metabolic pathway of doxylamine in the human on the basis of our data of the identified metabolites of doxylamine.

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Multi-Stakeholders in Public and Cultural Diplomacies as Seen through the Lens of Public-Private Partnerships: A Comparative Case Study of Germany and South Korea

  • Kim, Hwa Jung
    • Journal of Contemporary Eastern Asia
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    • v.17 no.1
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    • pp.68-93
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    • 2018
  • With the emergence of partnerships with private actors in public and cultural diplomacies, complexity in the relations among the various stakeholders involved has arisen, and yet the relevant research is insufficient to shed any beneficial light on such issues. By looking at public-private partnerships, the present study determined that resource dependence, trust, and risk are the main factors affecting the feasibility of partnerships, and inductively developed propositions on their effects. In an explorative case study, Germany (decentralized mode of governance) and South Korea (centralized mode of governance) were compared as exemplary contrasting system designs. The results revealed that risk and trust are likely to affect the feasibility of partnerships, whereas resource dependence is not. The following additional findings also were made: (1) there are cultural actors in a 'for profit, but with non-profit purposes' sector; (2) an interpersonal level of trust positively affects partnerships; (3) 'taking risks' brings about 'innovation'; (4) the existence of international commonalities between any two cases depending on the actors' shared role, whether public or private; (5) public actors' emphasis on mutual trust, program budget and execution, innovations coming from taking risks, commitment and ownership, and unexpected situations; (6) private actors' consideration of 'publicness' and grant-seeking or financial support as important incentives, and their desire that public actors to show more trust, professionalism, and ownership with less control over budget execution. With its qualitative approach and in-depth analysis, the present study yielded new insights, notwithstanding the relatively small sample data.

NMR study of the interaction of T4 Endonuclease V with DNA

  • Lee, Bong-Jin;Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.80-80
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    • 1995
  • T4 Endonuclease V (Mw 16,000) acts as a repair enzyme for UV induced pyrimidine dimers in DNA. Many researchers have studied the biochemical characteristics of the enzyme. However the precise action mechanism of T4 endo V has not fully elucidated yet. In our laboratory NMR spectroscopy technique is being used for the structural study of T4 endo V. Because of its low temperature stability and high content of ${\alpha}$-helix, the conventional $^1$H NMR technique was inapplicable. Therefore we utilized stable isotope labeling technique and so far prepared about 10 amino acid specific labeled proteins. The HSQC spectra of amino acid specific labeled proteins will help us to interpret the triple resonance 3D, 4D data which are under processing, We also studied the behaviors of specific amino acid residues whose roles might be critical. When the enzyme labeled by $\^$15/N-Thr was mixed with the substrate oligonucleotide (semispecific -TT- sequence), one crosspeak in its HSQC spectrum was completely desappeared, which means that one of seven Thr residues is in the binding site of the enzyme with DNA, This result is well consistent with previous report that implicated the Thr 2 residue in the activity of the enzyme. Similar studies were carried on the behaviors of Arg and Tyr residues.

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The individual influence factors of voluntary turnover among knowledge workers in China: A case study of Huawei

  • Feng, Hua;Cao, Mengyin
    • Journal of Contemporary Eastern Asia
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    • v.16 no.2
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    • pp.3-19
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    • 2017
  • "Knowledge worker" is a term used to describe highly educated people whose role in the workforce involves the use of information and knowledge. Knowledge workers are key to the success of a great many organizations. Many Chinese companies find that the voluntary turnover rate among their knowledge workers is high. The purpose of this paper is to explore, from the perspective of knowledge workers and the characteristics peculiar to them, the factors that influence this high voluntary turnover rate. The models used in this paper, that will enable understanding of the characteristics of knowledge workers in China, are the Chinese Personality Assessment Inventory and the Five Factors Model. In the case of Huawei, analysis using these models leads to the conclusion that the main factors that influence the voluntary turnover of knowledge workers in China are: age; gender; degree of extraversion and how open the person is to new experiences; family orientation; and occupation. This study leads to the conclusion that the company should pay closer attention to three groups: female employees who put a high value on their families; employees from non-R&D departments; and younger employees who show a high degree of openness to new experiences.

Backbone assignments of 1H, 15N and 13C resonances and secondary structure prediction of MRA1997 from Mycobacterium tuberculosis H37Rv

  • Kim, Hyojung;Kim, Yena;Lee, Ki-Young;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.1
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    • pp.49-53
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    • 2015
  • MRA1997 is a 76-residue conserved hypothetical protein of Mycobacterium tuberculosis H37Ra, one of the most pathogenic bacterial species and the causative agent of tuberculosis. In this study, the sequence-specific backbone resonance assignment of MRA1997 was performed using NMR spectroscopy. Approximately 88.3% of the total resonances could be unambiguously assigned. By analyzing deviations of the $C{\alpha}$ and $C{\beta}$ chemical shift values, the secondary structure of MRA1997 was calculated. The result revealed that secondary structure of MRA 1997 consists of one ${\alpha}$-helix and five ${\beta}$-sheets. Our structural study will be a footstone towards the characterization of the three-dimensional structure of MRA1997.

Backbone 1H, 15N, and 13C resonance assignments and secondary structure prediction of SAV2228 (translation initiation factor-1) from Staphylococcus aureus

  • Kim, Do-Hee;Jang, Sun-Bok;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.16 no.2
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    • pp.162-171
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    • 2012
  • SAV2228 has an OB (Oligomer-Binding)-motif which is frequently used for nucleic acid recognition. To characterize the activity of translation initiation factor-1 (IF-1) from Staphylococcus aureus, SAV2228 was expressed and purified in Escherichia coli. We acquired 3D NMR spectra showing well dispersed and homogeneous signals which allow us to assign 94.4% of all $^1HN$, $^{15}N$, $^{13}C{\alpha}$, $^{13}C{\beta}$ and $^{13}CO$ resonances. We could predict a secondary structure of SAV2228 using TALOS and CSI from NMR data. SAV2228 was consisted of one ${\alpha}$-helix and five ${\beta}$-sheets. The predicted secondary structure, ${\beta}-{\beta}-{\beta}-{\alpha}-{\beta}-{\beta}$, was similar to other bacterial IF-1, but it was not completely same to the eukaryotic one. Assigned NMR peaks and secondary structre prediction can be used for the study on interaction with nucleic acid in the future.

Crystallization and X-ray crystallographic analysis of the PH-like domain of lipid transfer protein anchored at membrane contact sites from Saccharomyces cerevisiae

  • Tong, Junsen;Im, Young Jun
    • Biodesign
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    • v.5 no.4
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    • pp.136-140
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    • 2017
  • Lam6 is a member of sterol-specific ${\underline{l}ipid$ transfer proteins ${\underline{a}}nchored$ at ${\underline{m}ebrane$ contact sites (LAMs). Lam6 localizes to the ER-mitochondria contact sites by its PH-like domain and the C-terminal transmembrane helix. Here, we purified and crystallized the Lam6 PH-like domain from Saccharomyces cerevisiae. To aid crystallization of the Lam6 PH-like domain, T4 lysozyme was fused to the N-terminus of the Lam6 PH-like domain with a short dipeptide linker, GlySer. The fusion protein was crystallized under the condition of 0.1 M HEPES-HCl pH 7.0, 10% (w/v) PEG 8000, and 0.1 M $Na_3$ Citrate at 293K. X-ray diffraction data of the crystals were collected to $2.4{\AA}$ resolution using synchrotron radiation. The crystals belong to the orthorhombic space group $P2_12_12_1$ with unit cell parameters $a=59.5{\AA}$, $b=60.1{\AA}$, and $c=105.6{\AA}$. The asymmetric unit contains one T4L-Lam6 molecule with a solvent content of 58.7%. The initial attempt to solve the structure by molecular replacement using the T4 lysozyme structure was successful.