• Applied Biological Chemistry
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• v.32 no.4
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• pp.393-400
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• 1989

The amounts of $^{14}CO_2$ evolved during the $^{14}C-bentazon$ aging in soil for 3 and 6 months were 6.1 and 14.8% of the original radioactivity, respectively. The presence of earthworms in soil tended to increase the uptake of $^{14}C-bentazon$ by the roots of rice plants, even if it was not statistically significant. The evolution of $^{14}CO_2$ from $^{14}C-bentazon$ in soil increased in the presence of rice plants and earthworms compared with in the absence of them. The uptake of $^{14}C-bentazon$ residues by rice plants decreased remarkably with increasing the aging period within the limit of 3 months both in the absence and presence of earthworms, but there is not much difference between 3-month-aging and 6-month-aging. Much larger amounts of $^{14}C-labelled$ compounds were translocated to the shoots, compared with the data from a previous investigation using maize plants. The amount of non-extractable bound residue increased remarkably with the aging period up to 3 months. The polarity of the compounds extracted from soil increased with the aging and the growing of rice plants, indicating the formation of some polar metabolites.

• Molecules and Cells
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• v.38 no.5
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• pp.409-415
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• 2015

Low-barrier hydrogen bonds (LBHBs) have been proposed to have important influences on the enormous reaction rate increases achieved by many enzymes. ${\Delta}^5$-3-ketosteroi isomerase (KSI) catalyzes the allylic isomerization of ${\Delta}^5$-3-ketosteroid to its conjugated ${\Delta}^4$-isomers at a rate that approache the diffusion limit. Tyr14, a catalytic residue of KSI, has been hypothesized to form an LBHB with the oxyanion of a dienolate steroid intermediate generated during the catalysis. The unusual chemical shift of a proton at 16.8 ppm in the nuclear magnetic resonance spectrum has been attributed to an LBHB between Tyr14 $O{\eta}$ and C3-O of equilenin an intermediate analogue, in the active site of D38N KSI. This shift in the spectrum was not observed in Y30F/Y55F/D38N and Y30F/Y55F/Y115F/D38N mutant KSIs when each mutant was complexed with equilenin, suggesting that Tyr14 could not form LBHB with the intermediate analogue in these mutant KSIs. The crystal structure of Y30F/Y55F/Y115F/D38N-equilenin complex revealed that the distance between Tyr14 $O{\eta}$ and C3-O of the bound steroi was within a direct hydrogen bond. The conversion of LBHB to an ordinary hydrogen bond in the mutant KSI reduced the binding affinity for the steroid inhibitors by a factor of 8.1-11. In addition, the absence of LBHB reduced the catalytic activity by only a factor of 1.7-2. These results suggest that the amount of stabilization energy of the reaction intermediate provided by LBHB is small compared with that provided by an ordinary hydrogen bond in KSI.

• 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.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.279-285
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• 1993

Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.

• Journal of Life Science
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• v.28 no.10
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• pp.1140-1146
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• 2018

Dual-specificity phosphatases (DUSPs) play a role in cell growth and differentiation by modulating mitogen-activated protein kinases. DUSPs are considered targets for drugs against cancers, diabetes, immune diseases, and neuronal diseases. Part of the DUSP family, DUSP19 modulates c-Jun N-terminal kinase activity and is involved in osteoarthritis pathogenesis. Here, we report screening of cavity-creating mutants and the crystal structure of a cavity-creating L75A mutant of DUSP19 which has significantly enhanced enzyme activity in comparison to the wild-type protein. The crystal structure reveals a well-formed cavity due to the absent Leu75 side chain and a rotation of the active site-bound sulfate ion. Despite the cavity creation, residues surrounding the cavity did not rearrange significantly. Instead, a tightened hydrophobic interaction by a remote tryptophan residue was observed, indicating that the protein folding of the L75A mutant is stabilized by global folding energy minimization, not by local rearrangements in the cavity region. Conformation of the rotated active site sulfate ion resembles that of the phosphor-tyrosine substrate, indicating that cavity creation induces an optimal active site conformation. The activity enhancement by an internal cavity and its structural information provide insight on allosteric modulation of DUSP19 activity and development of therapeutics.

• Bulletin of the Korean Chemical Society
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• v.31 no.6
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• pp.1519-1526
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• 2010

The behavior of peptide or protein solutes in saline aqueous solution is a fundamental topic in physical chemistry. Addition of ions can strongly alter the thermodynamic and physical properties of peptide molecules in solution. In order to study the effects of added ionic salts on protein conformation and dynamics, we have used the molecular dynamics (MD) simulations to investigate the behavior of Staphylococcus aureus Hfq protein under two different ionic concentrations: 0.1 M NaCl and 1.0 M NaCl in presence and absence of RNA (a hepta-oligoribonucleotide AU5G). Hfq, a global regulator of gene expression is highly conserved and abundant RNA-binding protein. It is already reported that in vivo the increase of ionic strength results in a drastic reduction of Hfq affinity for $Q{\beta}$ RNA and reduces the tendency of aggregation of Escherichia coli host factor hexamers. Our results revealed the crucial role of 0.1 M NaCl Hfq system on the bases with strong hydrogen bonding interactions and by stabilizing the aromatic stacking of Tyr42 residue of the adjacent subunits/monomers with the adenine and uridine nucleobases. An increase in RNA pore diameter and weakened compactness of the Hfq-RNA complex was clearly observed in 1.0 M NaCl Hfq system with bound RNA. Aggregation of monomers in Hfq and the interaction of Hfq with RNA are greatly affected due to the presence of high ionic strength. Higher the ionic concentration, weaker is the aggregation and interaction. Our results were compatible with the experimental data and this is the first theoretical report for the experimental study done in 1980 by Uhlenbeck group for the present system.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.50 no.1
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• pp.11-18
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• 2024

Recently, there have been attempts to claim the hair moisturizing effect for a hair care product, however there has not yet been an official evaluation method because heating temperature for hair has not been established. This study was conducted to establish a quantitative evaluation for hair water content. In order to observe the behavior of water inside hair, heat was applied to hair with various temperatures using thermogravimetric dry residue. As the heating temperature increased, the amount of moisture released from the hair increased. As a result of evaluating hair using a differential scanning calorimeter (DSC), a unique phenomenon in which a rapid endothermic reaction occurs around 75 ℃ was observed. This phenomenon was also observed in different ethnic hair. In hair that damaged the hair cuticle barrier with oxidation and heat, this rapidly rising endothermic reaction temperature occurred at 77 ℃, which was slightly higher, and 73 ℃ was observed when this hair was applied with polar oil, conditioning polymer, or keratin protein. To determine how this reaction affects the hair surface, friction test was performed using an atomic force microscope. When heated above 75 ℃, cuticle friction increased, however when heated above 90 ℃, there was no change in hair cuticle friction. Finally, it was confirmed that around 75 ℃ is the critical temperature at which desorption of water bound to the hair occurs. It is suggested that a heating temperature of 75 ℃ is the optimal temperature for detecting and quantifying the moisture content of hair, and that approximately 10% detected at 75 ℃ can be a standard value for hair moisture content.

• The Korean Journal of Pesticide Science
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• v.8 no.4
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• pp.299-308
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• 2004

In order to elucidate the behavior of bensulfuron-methyl, a sulfonylurea herbicide, in a soil/plant microecosystem, rice plants (Oryza sativa L.) were grown for 12 weeks in the specially made stainless steel pots (17cm I.D. $\times$ 10cm H.) containing two different paddy soils treated with fresh and 13-week-aged residues of [phenyl-$^{14}C$]bensulfuron-methyl, respectively. During the aging period, the mineralization to $^{14}CO_2$ from soil A (OM, 3.59%; CEC, 7.65 $cmol^+\;kg^{-1}$; texture, sandy clay loam) and B (OM, 1.62%; CEC, 4.51 $cmol^+\;kg^{-1}$; texture, sandy loam) amounted to 6.79 and 10.15% of the originally applied $[^{14}C]$bensulfuron-methyl, respectively. The amounts of $^{14}CO_2$ evolved from the soils with fresh residues were higher than those from the soils with aged residues. At harvest after 12-week growing, $^{14}C$-radioactivity absorbed and translocated into rice plants from soils A and B containing fresh residues of bensulfuron-methyl was 1.53 and 4.40%, while 4.04 and 6.37% in the two soils containing aged residues, respectively. Irrespective of aging and soil type, the $^{14}C$-radioactivity remaining in soil ranged from 80.41 to 98.87% of the originally applied $[^{14}C]$bensulfuron-methyl. The solvent extractability of tile soils was $39.25\sim70.39%$, showing the big differences among the treatments. Most of the nonextractable soil-bound residues of $[^{14}C]$bensulfuron-methyl were incorporated into the fulvic acid fraction$(61.32\sim76.45%)$. Comparing the microbial activity of the soils with rice plants grown with that of the soils without them, the former was $1.6\sim3.0$ times higher than the latter. However, it did not correlate with the $^{14}CO_2$ evolution.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.4
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• pp.247-256
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• 2013

In this study, alginate beads containing birnessite (Bir-AB), a highly reactive oxidative catalyst for the transformation of phenolic compounds, was prepared and its 1-naphthol (1-NP) removal efficiency was investigated in a batch test. Based on scanning electron microscopy image, it can be inferred that the alginate gel cluster acts as a bridge which bind the birnessite particles together. Kinetic experiment with Bir-AB of different mixing ratios of birnessite to alginate (Bir : AG=0.25 : 1~1 : 1 w/w) indicate that pseudo-first order kinetic constants, $k(hr^{-1})$ for the 1-NP removals increased about 1.5 times when the birnessite mixing ratio was doubled. The removals of 1-NP was found to be dependent on solution pH and the pesudo-first order rate constants were increased from 0.331 $hr^{-1}$ at pH 10 to 0.661 $hr^{-1}$ at pH 4. The analysis of total organic carbon for the reaction solutions showed that a higher removal of dissolved organic carbon was achieved with Bir-AB as compared to birnessite. HPLC chromatographic analysis of the methanol extract after reaction of 1-NP with Bir-AB suggest that the reaction products could be removed through incorporation into the aliginate beads as a bound residue. Mn ions produced from the oxidative transformation of 1-NP by birnessite were also removed by sorption to Bir-AB. The Bir-AB was recovered quantitatively by simple filtration and was reused twice without significant loss of the initial reactivity.