• 한국수산과학회지
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• 제38권5호
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• pp.286-290
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• 2005

Oxytocin (OT)-related peptide, isotocin was purified from the brain extract of conger eel (Conger myriaster) using reverse-phase, ion-exchange and size exclusion high performance liquid chromatography (HPLC). The sequence of the peptide, with a molecular weight of 967.30 Da, was determined as Cys-Tyr-Ile-Ser-Asn­Cys-Pro-Ile-Gly-$NH_2$, where the Cys between 1st and 6th residues made an intramolecular disulfide bridge by the automated amino acid sequence analysis and MALDI-TOF mass spectrometry. The sequence was confirmed by identical elution with the purified and synthetic peptide using the HPLC system. As a result of homology investigation, the primary structure of this peptide was the same as that of OT -superfamily member, isotocin. The synthetic peptide showed a contractile activity at a minimal effective concentration of $10^{-7}M$ on the intestinal smooth muscle of goldfish (Carassius auratus).

• 대한수의학회지
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• 제39권1호
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• pp.45-54
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• 1999

In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398 of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide synthesizer. The specificity and identification of the antisera were tested by analysis of transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR. MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as endoplasmic reticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity was abolished when anti-MOR sera were preincubated with the peptide against which they were raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which was determined to be within the last amino acid, 391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was observed in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain areas showed the mirrored pattern observed in auto radiographic studies of mu-opioid binding as well as a pattern similar to that seen by is situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future.

• 한국수산과학회지
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• 제38권1호
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• pp.6-11
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• 2005

Vasopressin (VP)-related peptide was purified from the brain extract of conger eel (Conger myriaster) by reverse-phase, ion-exchange high performance liquid chromatography (HPLC). This peptide with a molecular weight of 1,051.2 Da was determined as $H-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Arg-Gly-NH_2$, whose Cys residues made an intramolecular disulfide bridge by the automated amino acid sequence analysis, MALDI- TOF mass spectrometry. It's sequence was confirmed by identity of the elution position with the synthetic peptide in HPLC system. As a result of homology investigation, the primary structure of this peptide was the same as that of VP-superfamily member, $[Arg^8]-vasotocin$. The synthetic peptide showed a contractile activity at a minimal effective concentration of $10^{-10}\;M$ on the intestinal smooth muscle of goldfish.

• International Journal of Oral Biology
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• 제39권4호
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• pp.169-176
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• 2014

A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format $O_{(1-6)}XXXXX-NH_2$, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-$NH_2$ (pep-1) and WXXXXN-$NH_2$ (pep-2), and one positioned peptide, RRRWRN-$NH_2$ (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic ${\alpha}$-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

• Journal of Microbiology and Biotechnology
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• 제27권11호
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• pp.1999-2009
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• 2017

The secretion efficiency of a protein in a Sec-type secretion system is mainly determined by an N-terminal signal peptide and its combination with its cognate protein. Five signal peptides, namely, two synthetic Sec-type and three Bacillus licheniformis alpha-amylase-derived signal peptides, were compared for periplasmic expression of the human growth hormone (hGH) in E. coli. Based on in silico predictions on the signal peptides' cleavage efficiencies and their corresponding mRNA secondary structures, a number of amino acid substitutions and silent mutations were considered in the modified signal sequences. The two synthetic signal peptides, specifically designed for hGH secretion in E. coli, differ in their N-terminal positively charged residues and hydrophobic region lengths. According to the mRNA secondary structure predictions, combinations of the protein and each of the five signal sequences could lead to different outcomes, especially when accessibility of the initiator ATG and ribosome binding sites were considered. In the experimental stage, the two synthetic signal peptides displayed complete processing and resulted in efficient secretion of the mature hGH in periplasmic regions, as was demonstrated by protein analysis. The three alpha-amylase-derived signal peptides, however, were processed partially from their precursors. Therefore, to achieve efficient secretion of a protein in a heterologous system, designing a specific signal peptide by using a combined approach of optimizations of the mRNA secondary structure and the signal peptide H-domain and cleavage site is recommended.

• 한국자원식물학회지
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• 제30권1호
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• pp.1-7
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• 2017

본 연구는 항암 단백질로 알려진 lunasin peptide의 산업적 활용성을 높이고자 lunasin peptide를 생산할 수 있는 시스템을 개발하고, 생산된 lunasin peptide가 식물유래 lunasin peptide의 생리활성을 가지는지 chromatin binding 활성과 histone acetylation 억제활성을 통해 평가하였다. 그 결과 E. coli와 P. pastoris를활용하여 재조합 lunasin peptide를 생산했으며, 생산 된 재조합 lunasin 펩타이드가 식물유래 lunasin peptide의 chromatin binding 활성과 histone acetylation 억제활성을 나타냄을 확인할 수 있었다. 따라서, 본 실험 연구의결과를 토대로 lunasin 펩타이드의 대량생산이 진행된다면 천연물 유래 생리활성 물질로서 효과적이면서도 안전한 기능성 식품소재로의 산업적 활용이 가능할 것으로 기대된다.

• BMB Reports
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• 제44권10호
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• pp.669-673
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• 2011

Human methionine sulfoxide reductase B3A (hMsrB3A) is an endoplasmic reticulum (ER) reductase that catalyzes the stereospecific reduction of methionine-R-sulfoxide to methionine in proteins. In this work, we identified an antimicrobial peptide from hMsrB3A protein. The N-terminal ER-targeting signal peptide (amino acids 1-31) conferred an antimicrobial effect in Escherichia coli cells. Sequence and structural analyses showed that the overall positively charged ER signal peptide had an Argand Pro-rich region and a potential hydrophobic ${\alpha}$-helical segment that contains 4 cysteine residues. The potential ${\alpha}$-helical region was essential for the antimicrobial activity within E. coli cells. A synthetic peptide, comprised of 2-26 amino acids of the signal peptide, was effective at killing Gram-negative E. coli, Klebsiella pneumoniae, and Salmonella paratyphi, but had no bactericidal activity against Gram-positive Staphylococcus aureus.

• Restorative Dentistry and Endodontics
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• 제34권2호
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• pp.120-129
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• 2009

본 연구에서는 DSP (dentin sialoprotein)에서 유래된 합성 펩타이드를 동물실험 모델에 적용하여 치수노출 부위에서 상아질 재생을 확인하고, 기존 치 수복조제와의 성능 비교를 통해 새로운 치수복조제로서의 가능성을 확인하고자 하였다. 6마리 비글견의 73개의 치아를 이용하여, 실험적으로 치수를 노출하고 직접 치수 복조술을 시행하였다. 사용한 치수복조제는 (1) $Ca(OH)_2$ (CH군) (2) DSP유도 합성 펩타이드 (PEP군) (3) 합성 펩타이드와 $Ca(OH)_2$ 혼합제 (PEP+CH군) (4) White MTA (WMTA군) 이다. 노출된 치수에 치수복조제를 적용한 후 와동은 강화형 글라스 아이오노머로 충전하였다. 시술 후 2주, 1 개월 및 3개월에 각각 2마리씩 비글견을 희생시키고 조직시편을 제작하였다. 시편은 H&E 염색 후 광학 현미경으로 치수 염증 반응과 경조직 형성 정도를 관찰하였다. PEP군에서는 17개의 시편 중 3 개의 시편에서만 경조직 형성을 관찰할 수 있었으며, 대부분의 시편에서 적절한 치수 회복을 관찰할 수 없었다. PEP군은 CH군에 비해 심한 염증반응을 보이고, 경조직 형성은 불량하였다. CH군과 WMTA군은 기계적으로 노출된 치수에서의 염증반응과 경조직 형성에 있어서 유사한 결과를 보였다.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.276-281
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• 1999

In order to determine the functional region of the antifungal protein (AFP) isolated from Aspergillus giganteus responsible for growth inhibitory activity and the promotion of phospholipid vesicle aggregation, overlapping peptides covering the complete sequence of AFP were synthesized. The antibiotic activity against bacterial, fungal, and tumor cells, and the vesicle-aggregation activity of the synthetic peptides were investigated. The AFP functional sequence responsible for antibiotic and vesicle-aggregation activity was determined to be located within the region between AFP residues 19 to 32. AFP (19-32) exhibited an a-helical conformation in a cell membrane-like environment. AFP (19-32) displayed potent antibiotic activity against bacterial, fungal, and tumor cells without peptide toxicity as indicated by hemolysis. Accordingly, AFP (19-32) could be used as a good model for the design of effective antibiotic agents with powerful antibiotic activity yet without any cytotoxic effects against the host organism.

• Journal of Electrical Engineering and Technology
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• 제2권2호
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• pp.274-279
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• 2007

This study focuses on the enhancement of maskless photolithography as well as the peptide synthesis application with single crystalline silicon micromirrors. A single crystalline silicon micromirror array has been designed and fabricated in order to improve its application to the peptide synthesis. A micromirror rotates about ${\pm}\;9^{\circ}$ at the pull-in voltage, which can range from 90.7 V to 115.1 V. A $210\;{\mu}m-by-210\;{\mu}m$ micromirror device with $270\;{\mu}m$ mirror pitch meets the requirements of an adequately precise separation for peptide synthesis. Synthetic 16 by 16 peptide array corresponds to the same number of micromirrors. The large size of peptide pattern and the separation facilitate biochip experiments using fluorescence assay. The peptide pattern has been synthesized on the GPTS-PEG200 surface with BSA-blocking and thereupon the background was acetylated to reject non-specific bindings. Hence, an averaged slope at the pattern edge has been distinguishably improved in comparison to patterning results from an aluminum micromirror.