• BMB Reports
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• 제29권2호
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• pp.99-104
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• 1996

Site-directed substitutions were made to change the Ile91 of restriction endonuclease EcoRV to either Val, Ala or Gly to identify the role of Ile91 in recognition and catalysis, since substitution of Ile91 with Leu afforded dramatic effects on the activity and properties of restriction endonuclease EcoRV. These changes alter the size of the hydrophobic side chain at position 91 and thus might have revealed the reason for the altered phenotype of Ile91Leu. However, the properties of Ile91Val and Ile91Ala mutants were much like wild type EcoRV, in both activity and metal ion preference. Ile91Gly had very little activity with either $Mg^{2+}$ or $Mn^{2+}$ as cofactors. To try to understand the unusual $Mn^{2+}$ profile of the Ile91Leu mutant, two double mutants, Ile91Leu;Asp90Asn and Ile91Leu;Glu45Met were created. Both double mutants were seriously disabled by the second amino acid change. Ile91Leu;Glu45Met had some residual activity in the $Mn^{2+}$ reaction buffer, whereas the Ile91Leu;Asp90Asn displayed no detectable activity.

• BMB Reports
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• 제39권2호
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• pp.150-157
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• 2006

Native grass carp (Ctenopharygodon idellus) growth hormone, has 5 cysteine amino acid residues, forms two disulphide bridges in its mature form. Recombinant grass carp growth hormone, when over-expressed in E. coli, forms inclusion bodies. In vitro oxidative renaturation of guanidine-hydrochloride dissolved recombinant grass carp growth hormone was achieved by sequential dilution and stepwise dialysis at pH 8.5. The redox potential of the refolding cocktail was maintained by glutathione disulphide/glutathione couple. The oxidative refolded protein is heterogeneous, and contains multimers, oligomers and monomers. The presence of non-disulphide-bond-forming cysteine in recombinant grass carp growth hormone enhances intermolecular disulphide bond formation and also non-native intramolecular disulphide bond formation during protein folding. The non-disulphide-bond-forming cysteine was converted to serine by PCR-mediated site-directed mutagenesis. The resulting 4-cysteine grass carp growth hormone has improved in vitro oxidative refolding properties when studied by gel filtration and reverse phase chromatography. The refolded 4-cysteine form has less hydrophobic aggregate and has only one monomeric isoform. Both refolded 4-cysteine and 5-cystiene forms are active in radioreceptor binding assay.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2949-2953
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• 2012

The glutathione S transferase (GST) family is a major part of cellular defense mechanisms against endogenous and exogenous substances, many of which have carcinogenic potential. Alteration in the expression level or structure of the glutathione-S-transferase (GST) enzymes may lead to inadequate detoxification of potential carcinogens and consequently contribute to cancer development. A member of the glutathione-S-transferase (GST) family, GSTP1, is an attractive candidate for involvement in susceptibility to carcinogen-associated colorectal cancer. An $Ag{\rightarrow}G$ transition in exon 5 resulting in an Ile105Val amino acid substitution has been identified which alters catalytic efficiency. The present study investigated the possible impact of Ile105Val GSTP1 polymorphism on susceptibility to colorectal cancer. in Jordan We examined 90 tissue samples previously diagnosed with colorectal carcinoma, and 56 non-cancerous colon tissues. DNA was extracted from paraffin embedded tissues and the status of the GSTP1 polymorphism was determined using a polymerase chain reaction restriction fragment length polymorphism (RFLP) method. No statistically significant differences were found between colorectal cancer cases and controls for the GSTP1 Ile/Ile, Ile/Val and Val/Val genotypes. The glutathione S-transferase polymorphism was not associated with risk in colorectal cancer cases in Jordan stratified by age, sex, site, grade or tumor stage. In conclusion, the GSTP1 Ile105Val polymorphism is unlikely to affect the risk of colorectal cancer.

• 한국인쇄학회지
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• 제16권1호
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• pp.55-67
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• 1998

현재, 디지털 하드카피에 있어서 색변환방법으로 신경망에 의한 비선형변환 방법은 많은 연구가 되어지고 있으며, 색변환 시스템으로서의 유용성이 확인되고 있다. 그러나, 학습용 패치의 제작시 여러 가지 프린트 왜곡에 의해서 톤 재현의 범위가 좁아지게 되고, 전체적으로 불균일한 학습용 패치가 얻어지게 된다. 그러므로 신경망 학습의 허용오차 범위가 줄어들게 되러 CIEL*a*b 에서 CMYK로의 정확한 색변환이 어렵게 되고, 재현된 결과물이 원고 화상과 큰 색차를 가지게 된다. 본 논문에서는 이러한 문제점들을 해결하시 위해 시지각성특성에 기반을 둔 프린터의 톤 재현 범위 확장법을 이용하여 신경망의 학습용 패치를 제작하여 CIEL*a*b 에서 CMYK로의 비성형 색변환 방법을 제안하고, 제안한 방법의 유용성을 확인하고자 하였다.

• 한국자기공명학회논문지
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• 제21권3호
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• pp.78-84
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• 2017

Pseudin is a naturally occurring 24 amino-acid-residue antimicrobial peptide derived from the skin of paradoxical frog Pseud's paradoxa. It shows potency against the bacteria and antibiotic-resistant bacteria strain, but has high cytotoxicity against mammalian cell. In our previous study, substitution of $Pro^{11}$ for Gly (Ps-P) increased bacterial cell selectivity but decreased the antibacterial activity of pseudin. In this study, we designed pseudin analogue, Ps-4K-P with increased cationicity up to +7 in Ps-P by substituting Glu14, Gln10, Gln24, and Leu18 with Lys. Ps-4K-P showed improved potent antibacterial activity with high bacterial cell selectivity. We determined the tertiary structure of Ps-4K-P in the presence of DPC micelles by NMR spectroscopy and it has a hinge structure at $Pro^{11}$ followed by three turn helices from $Pro^{11}$ to $Val^{23}$ at the C-terminus. Amphipathicity with increased cationicity as well as helix-hinge-helix structural motif provided by introduction of a Pro at position $Gly^{11}$ are the crucial factors which confer antibacterial activity with bacterial cell selectivity to Ps-4K-P.

• Asian-Australasian Journal of Animal Sciences
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• 제6권2호
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• pp.253-258
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• 1993

The study on the nutritive value and the potential use of two kinds (local vs. import) of sesame meal (SSM) in layer diets as substitute for soybean meal (SBM) at 25, 50, and 75% was carried out. Three hundred and thirty six layers were kept individually on a battery cage and fed isonitrogenous diets, 16% crude protein (CP), for 252 days. The local and the imported SSM contained on air dry basis 35.7 and 36.3% CP, 24.7 and 9.1% ether extract (EE), respectively. Their amino acid contents are all lower than those reported by NRC (1984). The local material contained remarkably lower lysine (0.44 vs. 0.94), methionine + cystine (1.06 vs. 1.75) and threonine (0.52 vs. 1.22) than the imported meal. Imported SSM could be substituted for SBM as high as 50% in layer ration without statistically adverse effect on egg production, feed intake, body weight gain, average egg weight and egg size, while local SSM was inferior to the control in all aspects. However, when production performance of the groups fed either kind of SSM was compared, no significant difference was found at the two lower substitution levels.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1453-1458
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• 2006

PKR-like endoplasmic reticulum (ER) kinase (PERK) is a type I transmembrane ER-resident protein containing a cytoplasmic catalytic domain with a Ser/Thr kinase activity, which is most closely related to the eukaryotic translation initiation factor-$2{\alpha}$ ($eIF2{\alpha}$) kinase PKR involved in the antiviral defense pathway by interferon. We cloned and expressed the PERK C-terminal kinase domain (cPERK) in Escherichia coli. Like PERK activation in cells under ER stress, wild-type cPERK underwent autophosphorylation when overexpressed in E. coli, whereas the cPERK(K621M) with a methionine substitution for the lysine at amino acid 621 lost the autophosphorylation activity. The activated form cPERK which was purified to near homogeneity, formed an oligomer and was able to trans-phosphorylate specifically its cellular substrate $eIF2{\alpha}$. Two-dimensional phosphoamino acids analysis revealed that phosphorylation of cPERK occurs at the Ser and Thr residues. The functionally active recombinant cPERK, and its inactive mutant should be useful for the analysis of biochemical functions of PERK and for the determination of their three-dimensional structures.

• 한국가금학회지
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• 제10권1호
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• pp.30-34
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• 1983

브로일러 사료에 있어서 피혁분의 어분대치 가능성을 검토하기 위하여 브로일러용 초생추 (♀) 240수를 사용하여 6주간의 사양시험을 실시하였다. 양질의 어분과 대치한 피혁분의 사용수준은 0%, 1%, 2%, 4%, 6%이었으며 시험경과 얻어진 성적은 다음과 같다. 1. 본 시험에 사용된 피혁분의 단백질함량은 74.41%, 조회분함량 17.9%, 크롬함량 2.0%이었고 pepsin소화율은 85%이었다. 2. 증체량. 사료섭취량 및 사료효율은 피혁분 4%수준까지든 별 차이를 보이지 않았으나 6%수준에서는 현저히 (p＜0.05) 떨어지는 결과를 보였다. 이러한 결과는 브로일러 사료에서 피혁분을 어분과 대치하여 4%수준까지 사용가능함을 시사한다.

• 대한수의학회지
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• 제44권4호
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• pp.571-579
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• 2004

In order to obtain the genetic informations of the Korean isolates of porcine circovirus 2 (PCV2), nucleotide sequences of total genome of three isolates and open reading frame 2 (ORF2) of four isolates were determined and compared with those of other reference PCV2 isolates. Nucleotide sequences of 3 isolates showed over 99% homology with those of reference strain (GenBank accession no. AF027217). Point mutations were mainly determined on ORF2 regions but little on ORF1 regions. The patterns of pointmutated sites and nucleotide substitution on ORF2 regions were generally consistent between Korean isolates, and these mutated sites observed in Korean isolates were also relatively similar to those of foreign isolates. Phylogenetic analysis of nucleotide or amino acid sequences showed that there were minor branches consisting of three clusters; cluster of Korea, Canada and America, cluster of Spain and Taiwan, and the last cluster of French and China isolates. These results suggested that Korean PCV2s were probably originated from North America such as Canada or USA. The genetic informations obtained from this study could be useful for the research of diagnosis and pathogenecity of PCV2.

• Bulletin of the Korean Chemical Society
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• 제33권9호
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• pp.2883-2889
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• 2012

LL-37 is the only antimicrobial peptide (AMP) of the human cathelicidin family. In addition to potent antimicrobial activity, LL-37 is known to have the potential to inhibit lipolysaccharide (LPS)-induced endotoxic effects. To provide the stability to proteolytic digestion and increase prokaryotic selectivity and/or anti-endotoxic activity of two Lys/Trp-substituted 19-meric antimicrobial peptides (a4-W1 and a4-W2) designed from IG-19 (residues 13-31 of LL-37), we synthesized the diastereomeric peptides (a4-W1-D and a4-W2-D) with D-amino acid substitution at positions 3, 7, 10, 13 and 17 of a4-W1 and a4-W2, respectively and the enantiomeric peptides (a4-W1-E and a4-W2-E) composed D-amino acids. The diastereomeric peptides exhibited the best prokaryotic selectivity and effective protease stability, but no or less anti-endotoxic activity. In contrast, the enantiomeric peptides had not only prokaryotic selectivity and anti-endotoxic activity but also protease stability. Our results suggest that the hydrophobicity and ${\alpha}$-helicity of the peptide is important for anti-endotoxic activity. In particular, the enantiomeric peptides showed potent anti-endotoxic and LPS-neutralizing activities comparable to that of LL-37. Taken together, both a4-W1-E and a4-W2-E holds promise as a template for the development of peptide antibiotics for the treatment of endotoxic shock and sepsis.