• Analytical Science and Technology
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• v.11 no.1
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• pp.73-78
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• 1998

The reverse detection heteronuclear multiple quantum coherence, HMQC study of metcyano complex of horse myoglobin(MbCN) has provided the complete assignment of hyperfine shifted resonances of heme carbons attached with proton(s). The application of HMQC experiment to the paramagnetic low-spin MbCN gives clear $^1H$ and $^{13}C$ coherences for the paramagnetic amino acid residues as well as heme side chains, and can be extended to the low-spin paramagnetic hemoprotein derivative for the assignment of natural abundance $^{13}C$ resonances. This assignment strategy can avoid possible ambiguities that may result from the sole utilization of $^1H$ nuclear Overhauser effect for the assignment of heme $^1H$ signals resonating in the diamagnetic region. The resulting 2,4-vinyl ${\alpha}$-carbons and 7-propionate ${\beta}$-carbon follow anomalous anti-Curie behavior, and are indicative of incoplanarity with heme plane. Magnetic/electronic asymmetry of heme induced by proximal histidine(His) makes spread that the hyperfine shifted heme carbon resonances over the range of 250 ppm at $25^{\circ}C$. These heme carbon resonances would be the much more sensitive probe than those of proton resonances in analyzing the nature of heme electronic structure of myoglobin.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.5
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• pp.614-628
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• 2019

Objective: The inhibitory leukocyte immunoglobulin-like receptors (LILRBs) play an important role in innate immunity. The present study represents the first description of the cloning and structural and functional analysis of LILRB1 and LILRB3 isolated from two genetically disparate chicken lines. Methods: Chicken LILRB1-3 genes were identified by bioinformatics approach. Expression studies were performed by transfection, quantitative polymerase chain reaction. Signal transduction was analyzed by western blots, immunoprecipitation and flow cytometric. Cytokine levels were determined by enzyme-linked immunosorbent assay. Results: Amino acid homology and phylogenetic analyses showed that the homologies of LILRB1 and LILRB3 in the chicken line 6.3 to those proteins in the chicken line 7.2 ranged between 97%-99%, while homologies between chicken and mammal proteins ranged between 13%-19%, and 13%-69%, respectively. Our findings indicate that LILRB1 and LILRB3 subdivided into two groups based on the immunoreceptor tyrosine-based inhibitory motifs (ITIM) present in the transmembrane domain. Chicken line 6.3 has two ITIM motifs of the sequence LxYxxL and SxYxxV while line 7.2 has two ITIM motifs of the sequences LxYxxL and LxYxxV. These motifs bind to SHP-2 (protein tyrosine phosphatase, non-receptor type 11) that plays a regulatory role in immune functions. Moreover, our data indicate that LILRB1 and LILRB3 associated with and activated major histocompatibility complex (MHC) class I and ${\beta}2-microglobulin$ and induced the expression of transporters associated with antigen processing, which are essential for MHC class I antigen presentation. This suggests that LILRB1 and LILRB3 are transcriptional regulators, modulating the expression of components in the MHC class I pathway and thereby regulating immune responses. Furthermore, LILRB1 and LILRB3 activated Janus kinase2/tyrosine kinase 2 (JAK2/TYK2); signal transducer and activator of transcription1/3 (STAT1/3), and suppressor of cytokine signaling 1 genes expressed in Macrophage (HD11) cells, which induced Th1, Th2, and Th17 cytokines. Conclusion: These data indicate that LILRB1 and LILRB3 are innate immune receptors associated with SHP-2, MHC class I, ${\beta}2-microglobulin$, and they activate the Janus kinase/signal transducer and activator of transcription signaling pathway. Thus, our study provides novel insights into the regulation of immunity and immunopathology.

• Korean Journal of Poultry Science
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• v.33 no.2
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• pp.151-158
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• 2006

In order to study the effects of supplementary $Safmannan^(R)$(Beta-glucan & MOS complex) and $World-Labs^(R)$ (multiple probiotics) on the performance, nutrient availability, small intestinal microflora and immune response in broiler chicks, one thousand hatched broilers ($Ross^(R)$ were assigned to 4 treatments: control(basal diet), $BMD^(R),\;Safmannan^(R)\;and\;World-Labs^(R)$. There were no significant differences in the performance and in serum IgG, ND titre. However parameters of leukocytes and erythrocytes were significantly different among treatments (p<0.05). Leukocytes and RBC of $World-Labs^(R)\;and\;Safmannan^(R)$ were mostly lower than $BMD^(R)$ and control whereby MCH and MCHC of $World-Labs^(R)\;and\;Safmannan^(R)$ were higher than other treatments. The cfu of intestinal microflora had no significant differences among treatments. The $BMD^(R)$ treatment was higher than others in amino acid and crude fat availability and $World-Labs^(R)$ was higher than others in crude fiber availability. It was concluded that supplements used in the present experiment did not significantly affect the production parameters. However, significant impact on blood parameters, especially on leucocytes, may need further investigation.

• Journal of Life Science
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• v.28 no.4
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• pp.389-397
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• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Journal of Sericultural and Entomological Science
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• v.47 no.2
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• pp.68-77
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• 2005

A characteristic of subtropical zones region MK-T2 compares with an gaeryangppong, and the 9-10 schedule the times when a leaf blooms to are fast, and ratio that a branch edge by the colds becomes lean showed 5.7%, and a growth of the new branch which went out delivers 67.2 cm, mulberry loaves of the new branch which went out, and 18.6, a form of a leaf is the 1.10 that length of a leaf grew more a bit than width of a leaf up. Thickness of a leaf is $228.2{\mu}m$, and an area is more similar than gaeryangppong as $225.6cm^2$. in plant taxonomy, the hair whom the style exists short with 0.7 mm, and go to the pistil head inside so as to be rare is distributed, and belong to Dolichostylae Pubescentes. The new branch cutting which executed without remedy processes was independent of a thickness of a case branch, and the form and 100% root was said, and an gaeryangppong compared with the fact that 10% root went out of 15 mm ideal, and was excellent very, and looked, a root went out a root the soil and water, all showed a characteristic to go out at central of a branch bases at 45% ratio. Length was 24.6 mm, and were water rate 78.8%, and mulberry of MK-T2 was carrying together sweetness and acidity to pH 4.7 while, besides, arrival was 19.21 Brix%. A larva period and pupa ratio, cocoon thickness ratio are almost similar to gaeryangppong, or weight of one cocoon, cocoon thickness, 20,002 cocoon quantity shows some results to drop, and be soft of a leaf, and feed value certifications are comparatively top-ranking. As a result of having analyzed amino acid of the 3rd day of 5th silkworm larva which bred to MK-T2, a collation absorbing an gaeryangppong went, and looked, but compared with a collation in case of tests to eat MK-T2, and looked, and the lie collations were not detected a difference at Leu, but MK-T2 tests were detected mutual almost similar amino acid creation. medical efficacy of the 3rd day of 5th silkworm larva ethanol extract which bred to MK-T2 and black results, histologic a case did not appear at HE dyeing about the kidney organization which extracted form the rats which ate a silkworm ethanol extract and dyeing all chemical organization immunity, and one step protein revelation became lower with almost unidentified levels.

• Journal of the Korean Chemical Society
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• v.54 no.5
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• pp.541-550
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• 2010

A new $N_4O_3$ heptadentate ligand, N,N'-Bis(2-hydroxybenzyl)-1,3-bis[(2-aminoethyl)amino]-2-propanol(H-BAP 4HCl)was synthesized. The hydrochloric acid salts of Br-BAP 4HCl, Cl-BAP 4HCl, $CH_3O$-BAP 4HCl and $CH_3$-BAP 4HCl containing Br-, Cl-, H-, $CH_3O-$ and $CH_{3^-}$ groups at the para-site of the phenol group of the H-BAP were synthesized. The structures of the ligands were confirmed by C. H. N. atomic analysis and $^1H$ NMR, $^{13}C$ NMR, UV-visible and mass spectra. The elemental stepwise protonation constants(${logK_n}^H$) of the synthesized $N_4O_3$ ligands showed six steps of the proton dissociation. The orders of the overall dissociation constants($log{\beta}_p$) of the ligands were Br-BAP < Cl-BAP < H-BAP < $CH_3O$-BAP < $CH_3$-BAP. The orders agreed well with that of Hammett substituent constants($\sigma_p$). The calculated stability constants($logK_{ML}$) between the ligands and transition metal ions agreed well with the order of the overall proton dissociation constants of the ligands but they showed a reverse order in Hammestt substituent constants($\sigma_p$). The order of the stability constants between the transition metal ions with the ligands were Co(II) < Ni(II) < Cu(II) > Zn(II) > Cd(II) > Pb(II).

• Journal of Life Science
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• v.33 no.10
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• pp.828-834
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• 2023

The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

• Radiation Oncology Journal
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• v.17 no.2
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• pp.151-157
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• 1999

Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

• Journal of Life Science
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• v.30 no.3
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• pp.304-312
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• 2020

Agarase can be used in the field of basic science, as well as for production of agar-derived high-functional oligosaccharides and bioenergy production using algae. In 2012, we summarized the classification, origin, production, and applications of agar. In this paper, we briefly review the literature on the recombinant expression of agarases from 2012 to the present. Agarase genes originated from 19 genera, including Agarivorans, Flammeovirga, Pseudoalteromonas, Gayadomonas, Catenovulum, Microbulbifer, Cellulophaga, Saccharophagus, Simiduia, and Vibrio. Of the 47 recombinant agarases, there were only two α-agarases, while the rest were β-agarases. All α-agarases produced agarotetraose, while β-agarases yielded many neoagarooligosaccharides ranging from neoagarobiose to neoagarododecaose. The optimum temperature ranged between 25 and 60℃, and the optimum pH ranged from 3.0 to 8.5. There were 14 agarases with an optimum temperature of 50℃ or higher, where agar is in sol state after melting. Artificial mutations, including manipulation of carbohydrate-binding modules (CBM), increased thermostability and simultaneously raised the optimum temperature and activity. Many hosts and secretion signals or riboswitches have been used for recombinant expression. In addition to gene recombination based on the amino acid sequence after agarase purification, recombinant expression of the putative agarase genes after genome sequencing and metagenome-derived agarases have been studied. This study is expected to be actively used in the application fields of agarase and agarase itself.

• Journal of Periodontal and Implant Science
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• v.37 no.3
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• pp.497-509
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• 2007

Bone morphogenetic proteins(BMPs) are regarded as members of the transforming growth $factor-{\beta}$ superfamily with characteristic features in their amino acid sequences. A number of studies have demonstrated the biologic activities of BMPs, which include the induction of cartilage and bone formation. Recently there was a attempt to overcome a limitation of mass production, and economical efficieny of rh-BMPs. The method producing PTD by using bacteria have advantages of acquiry a mass of proteins. Hences, a new treatment which deliver protein employed by protein transduction domain(PTD) has been tried. The purpose of this study was to evaluate the bone regenerative effect of TATBMP-2 and TAT-HA2-BMP-2 employed by PTD from HlV-1 TAT protein for protein translocation in the rat calvarial model. An 8mm calvarial, critical size osteotomy defect was created in each of 32 male Spraque-Dawley rats(weight $250{\sim}300g$). The animals were divided into 4 groups of 32 animals each (4 animals/group/healing interval). The defect was treated with TATBMP-2/ACS(Absorbable collagen sponge) (TATBMP-2 0.1mg/ml), TAT-HA2-BMP-2/ACS(TAT-HA2-BMP-2 0.1mg/ml), ACS alone or left untreated for surgical control(negative control). The rats were sacrificed at 2 or 8 weeks postsurgery, and the results were evaluated histologically. The results were as follows: New bone formation were not significantly greater in the TATBMP-2/ACS group relative to negative, and positive control groups. New bone was evident at the defect sites in TAT-HA2-BMP-2/ACS group relative to negative, positive control and TATBMP-2 groups. There were a little bone regeneration in TATBMP-2 groups. While, enhanced local bone formation were observed in TAT-HA2-BMP-2 group. But, The results was not the same in all rat defects. Therefore, further investigations are required to develop a method. which disperse homogenously, and adhere to target cells.