• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.86-86
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• 1997

Human annexin I is a member of annexin family of calcium dependent phospholipid binding proteins, which have been implicated in various physiological roles including phospholipase A$_2$ (PLA$_2$) inhibition, membrane fusion and calcium channel activity. In this work, the structure of N-terminally truncated human annexin I (Δ-annexin I) and its interactions with Ca$\^$2+/, ATP and cAMP were studied at atomic level by using $^1$H, $\^$15/N, $\^$l3/C NMR (nuclear magnetic resonance) spectroscopy. The effect of Ca$\^$2+/ binding on the structure of Δ-annexin I was investigated, and compared with that of Mg$\^$2+/ binding. The addition of Ca$\^$2+/ to Δ-annexin I caused some changes in the high field and low field regions of $^1$H NMR spectra. Whereas, upon addition of Mg$\^$2+/ to Δ-annexin I, almost no change could be observed. Also we found that the binding ratio of ATP to Δ-annexin I is 1. Because Δ-annexin I is a large protein with 35 kDa molecular weight, site-specific (carbonyl-$\^$l3/C, amide-$\^$15/N) labeling technique was used to determine the interaction sites of Δ-annexin I with Ca$\^$2+/ and ATP. Assignments of all the histidinyl carbonyl carbon resonances have been completed by using Δ-annexin I along with its specific 1,2-subdomain. The carbonyl carbon resonances originating from His52 and His246 of Δ-annexin I were significantly affected by Ca$\^$2+/ binding, and some Tyr and Phe resonances were also affected. The carbonyl carbon resonances originating from His52 is significantly affected by ATP binding, therefore His52 seems to be involved in the ATP binding site of Δ-annexin I.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.297-302
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• 1988

The protoplasts of Saccharomyces cerevisiae D-71, a thermophilic strain and Zygosaccharomyces rouxii SR-S, an osmotolerant strain were fused, and a fusant (FS-RN 1) was selected, then was characterized for its genetic stability, DNA content, cell capacity, growth rate, tolerance to salts and chemicals, $\beta$-fructofuranosidase level and ethanol fermenting activity. After 6 months of preservation, 5.8% of the fusant clones were segregated to parental types. The DNA content and cell capacity of the fusant were greater than those of the parental strains. Lag period of growth for the fusant was longer than those for the parents. The fusant colonies showed pink-color reaction to triphenyltetrazolium chloride(TTC) test. The fusant appeared to have resistances to NaCl at moderate levels between both parental strains, and resistances to KCI, sodium propionate and cycloheximide similar to either one of the parents. $\beta$-Fructofuranosidase activity of the fusant was 24.2$\times$10$^{-3}$/IU/g(dry wt) for 3 days culture. Ethanol yields ofter 4 days of fermentation by the fusant at 3$0^{\circ}C$ were : 6.0%(v/v) from 40% of glucose and 8.8%(v/v) from 20% of sucrose.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.102-108
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• 1994

Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

• Journal of the Korean Magnetic Resonance Society
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• v.2 no.2
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• pp.141-151
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• 1998

Human annexin I is a member of annexin family of calcium dependent phospholipid binding proteins, which have been implicated in various physiological roles including phospholipase A2(PLA2) inhibition, membrane fusion and calcium channel activity. In this work, the structure of N-terminally truncated human annexin I ({{{{ DELTA }}-annexin I) and its interactions with Ca2+, ATP and cAMP were studied at atomic level by using nuclear magnetic resonance (NMR) spectroscopy. The effect of Ca2+ binding on the structure of {{{{ DELTA }}-annexin I was investigated. The addition of Ca2+ to {{{{ DELTA }}-annexin I caused some changes in 13C NMR spectra. Carbonyl carbon resonances of some histidines were significantly broadened by Ca2+ binding. However, in the case of methionine, phenylalanine, and tyrosin, small changes could be observed. We found that ATP and cAMP bind {{{{ DELTA }}-annexin I, and the binding ratio of ATP to {{{{ DELTA }}-annexin I is 1. These results are well consistent with the report that cAMP and ATP interact with annexin I, and affect the calcium channels formed by annexin I. Because {{{{ DELTA }}-annexin I is a large protein with 35 kDa molecular weight, site-specific (carbonyl-13C) labeling technique was used to study the interaction sites of {{{{ DELTA }}-annexin I with Ca2+. NMR study was focused on the carbonyl carbon resonances of tyrosine, phenylalanine, methionine and histidine residues of {{{{ DELTA }}-annexin I because the number of these amino acids is small in the amino acid sequence of {{{{ DELTA }}-annexin I.

• The Transactions of the Korean Institute of Electrical Engineers C
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• v.53 no.1
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• pp.8-14
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• 2004

An O-mode Ultrashort Pulse Reflectometry (USPR) system has been designed and developed for the measurement of electron density profiles on the Sustained Spheromak Physics Experiment (SSPX) spheromak. In the original design of SSPX, peak densities were envisioned to be in the range of 0.5-3${\times}$10$^{14}$ cm$^{-3}$ . The total duration of formation and sustained discharges is typically ∼2 msec. Moreover, diagnostic access on SSPX is severely restricted. Such high density and short duration plasmas coupled with stringent diagnostic access are quite challenging for conventional reflectometer systems. In USPR, the SSPX diagnostic requirements have been successfully satisfied by employing up-converting mixers and monostatic horn/waveguide configuration. As a result, the USPR system has proven its applicability for the density measurement of a future fusion device. In the density profile measurements, the USPR system is capable of routinely generating density profiles with a temporal resolution of 57 $\mu$s. This paper presents details regarding the USPR fundamental principles, associated subsystems and laboratory tests as well as the experimental results obtained on SSPX

• Molecules and Cells
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• v.19 no.2
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• pp.191-197
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• 2005

The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.

• Journal of Korean Neurosurgical Society
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• v.61 no.5
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• pp.574-581
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• 2018

Objective : The zero-profile implant (Zero-P) is accepted for use in anterior cervical fusion for the treatment of degenerative cervical disease. However, evidence pertaining to its efficiency and safety in traumatic cervical injury is largely insufficient. This study aimed to compare the overall outcomes of patients who underwent Zero-P for traumatic cervical disc injury. Methods : Data from a total of 53 consecutive patients who underwent surgery for traumatic or degenerative cervical disc disease using the Zero-P were reviewed. Seventeen patients (group A) had traumatic cervical disc injury and the remaining 36 (group B) had degenerative cervical disc herniation. The fusion and subsidence rates and Cobb angle were measured retrospectively from plain radiographs. The patients' clinical outcomes were evaluated using the Japanese Orthopedic Association (JOA) score and Odom's criteria. Results : The C2-7 Cobb and operative segmental angles increased by $3.45{\pm}7.61$ and $2.94{\pm}4.59$ in group A; and $2.46{\pm}7.31$ and $2.88{\pm}5.49$ in group B over 12 postoperative months, respectively. The subsidence and fusion rate was 35.0% and 95.0% in group A; and 36.6% and 95.1% in group B, respectively. None of the parameters differed significantly between groups. The clinical outcomes were similar in both groups in terms of increasing the JOA score and producing a grade higher than "good" using Odom's criteria. Conclusion : The application of Zero-P in patients with traumatic cervical disc injury was found to be acceptable when compared with the clinical and radiological outcomes of degenerative cervical spondylosis.

• Applied Biological Chemistry
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• v.28 no.1
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• pp.13-18
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• 1985

To investigate the fusion of bacterial protoplasts, two auxotrophic mutants of Bacillus subtilis were isolated after treatment with nitrosoguanidine. Two auxotrophs were uracil-auxotroph and tryptophan-auxotroph. Back mutation frequencies of $ura^-$ and $trp^-$ -auxotrophs were $2.4{\times}10^{-8}$ and less than $1.0{\times}10^{-8}$, respectively. The optimal pH and temperature of protoplast formation with lysozyme were 6.5 and $30^{\circ}C$. The optimal lysozyme concentration was $200{\mu}g/ml$. The regeneration frequency of protoplast was 3.3%.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.160-166
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• 2008

A DNA fragment containing 2,079 base pairs from Bacillus circulans CGMCC 1416 was cloned using degenerate PCR and inverse PCR. An open reading frame containing 981 bp was identified that encoding 326 amino acids residues, including a putative signal peptide of 31 residues. The deduced amino acid sequence showed the highest identity (68.1%) with $endo-{\beta}-1,4-D-mannanase$ from Bacillus circulans strain K-1 of the glycoside hydrolase family 5 (GH5). The sequence encoding the mature protein was cloned into the pET-22b(+) vector and expressed in Escherichia coli as a recombinant fusion protein containing an N-terminal hexahistidine sequence. The fusion protein was purified by $Ni^{2+}$ affinity chromatography and its hexahistidine tag cleaved to yield a 31-kDa ${\beta}$-mannanase having a specific activity of 481.55U/mg. The optimal activity of the purified protein, MANB48, was at $58^{\circ}C$ and pH 7.6. The hydrolysis product on substrate locust bean gum included a monosaccharide and mainly oligosaccharides. The recombinant MANB48 may be of potential use in the feed industry.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.350-355
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• 2010

Among human antimicrobial peptides (hAMPs), DCD-1L has a broad spectrum of antimicrobial activity over a wide pH range and in high salt concentrations. It offers a promising alternative to conventional antibiotics. The 458-bp-long dermcidin cDNA was amplified by PCR using a human fetal cDNA library as a template. The 147-bp fragment of the MDCD-1L gene encoding an additional methionine residue was subcloned into the pTYB11 vector. Recombinant MDCD-1L was expressed as an intein fusion protein in E. coli, and then purified by affinity chromatography using chitin beads. A small peptide with a molecular mass of about 5 kDa was detected by tricine gel electrophoresis. The recombinant MDCD-1L peptide was purified from the gel and its amino acid sequence was determined by nanoLC-ESI-MS/MS analysis. The initiating amino acid, methionine, remained attached to the N-terminal region of recombinant MDCD-1L. Purified MDCD-1L showed antimicrobial activity against a Micrococcus luteus test strain.