• Toxicological Research
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• 제17권
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• pp.109-115
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• 2001

Abrus agglutinin was purified from the kernels of Abrus precatorius by Sepharose 4B affinity column chromatography followed by Sephadex G-100 gel filtration column chromatography. About 1.25 g of abrus agglutinin was obtained from 1 kg of the kernels. The LD$_{50}$ of abrus agglutinin is 5 mg/kg of body weight, which is less toxic than that of abrin, 20$\mu\textrm{g}$/kg body weight. The amino acid sequence of abrus agglutinin was determined by protein sequencing techniques and deduced from the nucleotide sequence of a cDNA clone encoding full length of abrus agglutinin. There are 258 residues, 2 residues and 267 residues in the A-chain, the linker peptide and the B-chain of abrus agglutinin, respectively. Abrus agglutinin had high homology to abrin-a (77.8%). The 13 amino acid residues involved in catalytic function, which are highly conserved among abrin and ricin, were also conserved within abrus agglutinin. The protein synthesis inhibitory activity of abrus agglutinin ($IC_{50}$/ = 3.5 nM) was weaker than that of abrin-a (0.05 nM). By molecular modeling followed by site-directed mutagenesis showed that Pro199 of abrus agglutinin A-chain located in amphipathic helix H and corresponding to Asn200 of abrin A-chain, can induce bending of helix H. This bending would presumably affect the binding of abrus agglutinin A-chain to its target sequence GpApGpAp, in the tetraloop structure of 285 r-RNA subunit and this could be one of major factors contributing to the relatively weak protein synthesis inhibitory activity and toxicity of abrus agglutinin.n.

• BMB Reports
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• 제37권2호
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• pp.254-259
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• 2004

The wild-type and temperature-sensitive (ts) repressor genes were cloned from the temperate mycobacteriophage L1 and its mutant L1cIts391, respectively. A sequencing analysis revealed that the $131^{st}$ proline residue of the wild-type repressor was changed to leucine in the ts mutant repressor. The 100% identity that was discovered between the two DNA regions of phages L1 and L5, carrying the same sets of genes including their repressor genes, strengthened the speculation that L1 is a minor variant of phage L5 or vice versa. A comparative analysis of the repressor proteins of different mycobacteriophages suggests that the mycobacteriophage-specific repressor proteins constitute a new family of repressors, which were possibly evolved from a common ancestor. Alignment of the mycobacteriophage-specific repressor proteins showed at least 7 blocks (designated I-VII) that carried 3-8 identical amino acid residues. The amino acid residues of blocks V, VI, and some residues downstream to block VI are crucial for the function of the L1 (or L5) repressor. Blocks I and II possibly form the turn and helix 2 regions of the HTH motif of the repressor. Block IV in the L1 repressor is part of the most charged region encompassing amino acid residues 72-92, which flanks the putative N-terminal basic (residues 1-71) and C-terminal acidic (residues 93-183) domains of L1 repressor.

• Molecules and Cells
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• 제42권1호
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• pp.56-66
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• 2019

Histidine triad nucleotide-binding protein (HINT) is a member of the histidine triad (HIT) superfamily, which has hydrolase activity owing to a histidine triad motif. The HIT superfamily can be divided to five classes with functions in galactose metabolism, DNA repair, and tumor suppression. HINTs are highly conserved from archaea to humans and function as tumor suppressors, translation regulators, and neuropathy inhibitors. Although the structures of HINT proteins from various species have been reported, limited structural information is available for fungal species. Here, to elucidate the structural features and functional diversity of HINTs, we determined the crystal structure of HINT from the pathogenic fungus Candida albicans (CaHINT) in complex with zinc ions at a resolution of $2.5{\AA}$. Based on structural comparisons, the monomer of CaHINT overlaid best with HINT protein from the protozoal species Leishmania major. Additionally, structural comparisons with human HINT revealed an additional helix at the C-terminus of CaHINT. Interestingly, the extended C-terminal helix interacted with the N-terminal loop (${\alpha}1-{\beta}1$) and with the ${\alpha}3$ helix, which appeared to stabilize the dimerization of CaHINT. In the C-terminal region, structural and sequence comparisons showed strong relationships among 19 diverse species from archea to humans, suggesting early separation in the course of evolution. Further studies are required to address the functional significance of variations in the C-terminal region. This structural analysis of CaHINT provided important insights into the molecular aspects of evolution within the HIT superfamily.

• 약학회지
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• 제36권3호
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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.

• 생물환경조절학회지
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• 제21권4호
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• pp.485-490
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• 2012

질소처리에 따른 여러 관엽식물의 생육반응 및 엽내 질소 함량을 구명하여 초기 생육을 단축시키고 관엽식물의 영양관리에 있어 기초자료를 제시하고자 하였다. 실험재료로 벤자민고무나무, 관음죽, 필로덴드론, 스파티필럼, 아이비 등 많이 이용되고 있는 관엽식물들을 선택하였다. 화분에 식재한 관엽식물에 질소량을 각각 120, 150, 180, $210mg{\cdot}L^{-1}$로 처리하고, 8주 후 식물의 초장, 폭, 엽수, 엽면적, 생체중 및 건물중 등을 측정하였다. 식물 부위별 질소 함량도 분석하였다. 벤자민고무나무와 아이비는 N1 처리에서 식물의 생육이 우수하고, 초장 및 엽수의 증가량이 가장 큰 것으로 나타났다. 필로덴드론과 관음죽은 N4 처리의 경우 식물 생육이 일부 향상되었으나, 초장 및 엽수의 증가량은 다른 질소수준들간에 큰 차이는 없었다. 특히 관음죽은 식물의 생장량이 거의 없었다. 스파티필럼은 N4 처리에서 식물의 생육이 우수하고, 초장 및 엽수의 증가량도 가장 큰 것으로 보여졌다. 각 식물별 적합한 질소수준이 다른 것으로 판단되었다. 질소수준별 처리가 엽록소 함량에 크게 영향을 주지 않았고, 식물 부위별 질소함량도 일관된 경향을 보여지지 않았다. 그러므로, 관엽식물에서는 잎 등 식물체 부위별 영양 상태보다 식물의 생육반응으로 영양상태를 진단하는 것이 더 적합한 것으로 판단되었다.

• BMB Reports
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• 제44권2호
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• pp.118-122
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• 2011

Most widely used secondary structure assignment methods such as DSSP identify structural elements based on N-H and C=O hydrogen bonding patterns from X-ray or NMR-determined coordinates. Secondary structure assignment algorithms using limited $C{\alpha}$ information have been under development as well, but their accuracy is only ~80% compared to DSSP. We have hereby developed SABA (Secondary Structure Assignment Program Based on only Alpha Carbons) with ~90% accuracy. SABA defines a novel geometrical parameter, termed a pseudo center, which is the midpoint of two continuous $C{\alpha}s$. SABA is capable of identifying $\alpha$-helices, $3_{10}$-helices, and $\beta$-strands with high accuracy by using cut-off criteria on distances and dihedral angles between two or more pseudo centers. In addition to assigning secondary structures to $C{\alpha}$-only structures, algorithms using limited $C{\alpha}$ information with high accuracy have the potential to enhance the speed of calculations for high capacity structure comparison.

• Steel and Composite Structures
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• 제21권3호
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• pp.573-588
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• 2016

Carbon Fiber Reinforced Plastic (CFRP) and Titanium Alloy (Ti6Al4V) stack, extensively used in aerospace structural components are assembled by fasteners and the holes are made using drilling process. Drilling of stack in one shot is a complicated process due to dissimilarity in the material properties. It is vital to have optimal machining condition and tool geometry for better hole quality and tool life. In this study the tool wear and hole quality were analysed by experimental analysis using three modified twist drills and online tool condition monitoring using Acoustics Emission (AE) sensor. Helix angle and point angle influence tool performance and cutting force. It was found that a tool geometry (TG1) with high helix angle of $35^{\circ}$ with low point angle $130^{\circ}$ results in reduction in thrust force of 150-500 N range but the TG2 also perform almost similar to TG1, but when compared with the AErms voltage generated during drilling it was found that progressive rise in voltage in TG1 is less with respect to TG2 which can be attributed to tool life. In process wear monitoring was done using crest factor as monitoring index. AErms voltage were measured and correlated with the performance of the drills.

• Journal of Microbiology
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• 제45권3호
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• pp.193-198
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• 2007

A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1441-1447
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• 2006

The light perception and phototransformation of phytochromes are the first process of the phytochrome-mediated light signal transduction. The chromophore ligation and its photochromism of various site-specific and deletion mutants of pea phytochrome A and bacterial phytochrome-like protein (Cph1) were analyzed in vitro. Serial truncation mutants from the N-terminus and C-terminus indicated that the minimal N-terminal domain for the chromophore ligation spans from the residue 78 to 399 of pea phytochrome A. Site-specific mutants indicated that several residues are critical for the chromophore ligation and/or photochromism. Histidine-324 appears to serve as an anchimeric residue for photochromism through its H-bonding function. Isoleucine-80 and arginine-383 playa critical role for the chromophore ligation and photochromism. Arginine-383 is presumably involved in the stabilization of the Pfr form of pea phytochrome A. Apparently, the amphiphilic ${\alpha}$-helix centered around the residue-391 is in the chromophore pocket and critical for the chromophore ligation.

• 한국잠사곤충학회지
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• 제41권1호
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• pp.54-60
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• 1999

염산 또는 수산화나트륨 수용액의 농도, 온도, 시간 등 처리조건에 따른 작잠견사의 가수분해율을 측정하였으며 산처리 작잠분말과 알칼리처리 작잠분말의 형태 및 구조특성을 주사전자현미경, 적외선 분광분석, X-선 회절분석 등의 방법으로 분석한 결과 다음과 같은 결과를 얻었다. 1. 작잠견사에 대한 염산 및 수산화나트륨의 처리농도와 온도가 높을수록 또한 처리시간이 길어질수록 작잠견사의 가수분해율은 증가하는 경향을 나타내었으나 고온, 고농도의 조건하에서 수산화나트륨을 작잠견사에 처리하는 경우 작잠견사의 가수분해율이 오히려 떨어지는 경향을 보였다. 2. 산 및 알칼리 처리에 의하여 제조된 작잠분말의 형태는 용해도가 30 wt% 부근에서는 섬유상의 형태를 유지하였으며 산처리 분말의 경우 용해도가 증가함에 따라 섬유축 방향으로 조각난 미세한 입자형태를 나타내었으나 알칼리처리 분말의 경우에는 부정형의 입자형태를 나타내었다. 3. 적외선 분광분석 결과 작잠분말은 처리조건과 무관하게 ${\beta}$-sheet 구조의 특성을 반영하는 1635, 1532, 1226, 960, 700 cm$^{-1}$ 흡수대와 ${\alpha}$-helix 구조의 특성을 반영하는 616 cm$^{-1}$ 흡수대가 뚜렷하게 나타나 작잠분말의 주요 conformation은 ${\beta}$-sheet 구조와 ${\alpha}$-helix 구조임을 확인하였다. 4. X-선 회절분석 결과 가수분해율이 낮을 경우 (35 wt%)에는 ${\beta}$-sheet 구조와 ${\alpha}$-helix 구조에 기인하는 회절 피크가 뚜렷하게 나타났으나 가수분해율이 80 wt% 이상으로 증가하면 2${\theta}$=17.1$^{\circ}$ 및 20.0$^{\circ}$의 회절피크만 강하게 나타났다.