• YAKHAK HOEJI
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• v.35 no.6
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• pp.473-482
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• 1991

Protein methylase II (S-adenosyl-L-methionine:protein carboxyl-O-methyltransferase; EC 2.1.1.24., PM II) was purified from chicken pancreas by subcellular fractionation, DEAE-cellulose chromatography, QAE-Sephadex A-50 chromatography, Sephadex G-75 chromatography, and Sephadex G-75 rechromatography. The purified PM II gave a single band upon polyarcrylamide gel electrophoresis both in the presence of SDS and in Tris glycine buffer without SDS. The pI value of purified PM II was identified as 5.7 on isoelectric focusing gel. Properties and activities of PM II were studied and the following results were obtained. 1) PM II from chicken pancreas was purified approximately 221-fold with a yield of 1.3%. 2) The purified PM II appear constituted of a single polypeptide chain of a molecular weight 46,800 daltons. 3) Hemoglobin exhibited the highest of methyl-accepting activity among the substrates tested. 4) The purified PM II has a $K_m$ of $4.67{\times}10^{-6}M$ and a $V_{max}$ of 37.5 pmoles of $methyl-^{14}C/min./mg$ enzyme for $SAM^{-14}CH_3$ as methyl donor in the presence of histone type II-As. 5) It is found that S-adenosyl-L-homocysteine is a competitive inhibitor for PM II with $K_i$ value of $3.23{\times}10^{-5}M$.

• Korean journal of applied entomology
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• v.28 no.3
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• pp.105-112
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• 1989

The nuclear polyhedrosis viruses of Bombyx mori (BmNPV) and Hyphantria cunea (HcNPV) were characterized by electron microscopic observation, SDS-PAGE of polyhedral and virion proteins, and restriction endonuclease analysis of viral DNAs. Polyhedra of BmNPV were octadecahedral in shape with the diameter of $3 \mu\textrm{m}$, whereas those of HcNPV showed irregular appearances having the diameter of $1.5~2\mu\textrm{m}$. Under alkaline protease inactivated condition, polyhedral proteins of two NPVs were resolved into a major polypeptide, 30~31 KD, and several minor polypeptides by SDS-PAGE. Examination of virion proteins by silver staining after SDS-PAGE showed that BmNPV was composed of 47 polypeptides with M.W. range of 9.6~112 KD and HcNPV was composed of 48 polypeptides with M.W. range of 9.4~111 KD. The approximate genome size of two NPVs were determined by restriction endonuclease analysis: BmNPV and HcNPV were 114.6 Kb, respectively.

• Genomics & Informatics
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• v.3 no.2
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• pp.61-65
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• 2005

Comparing 231 genes on chimpanzee chromosome 22 with their orthologous on human chromosome 21, we have found that 15 orthologs have indels within their coding sequences. It was rather surprising that significant number of genes have changed by indel, despite the shorter time since their divergence and led us hypothesize that indels and structural changes may represent one of the major mechanism of proteome evolution in the higher primates. Human T-complex protein 10 like (TCP 10L) is a representative having indel within its coding sequence. Gene structure of human TCP10L compared with chimpanzee TCP10L gene showed 16 base pair difference in genomic DNA. As a result of the indel, frame shift mutation occurs in coding sequence (CDS) and human TCP10L express longer polypeptide of 21 amino acid residues than that of chimpanzee. Our prediction found that the indel may affect to dramatic change of secondary protein structure between human and chimpanzee TCP10L. Especially, the structural changes in the C-terminal region of TCP10L protein may affect on the interacting potential to other proteins rather than DNA binding function of the protein. Through these changes, TCP10L might influence gene expression profiles in liver and testis and subsequently influence the physiological changes required in primate evolution.

• The Journal of Dong Guk Oriental Medicine
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• v.8 no.1
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• pp.117-131
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• 1999

Yangkyuksanhoa-tang is frequently used for cerebrovascular accident(CVA). The present study was performed to investigate the effect of Yangkyuksanhoa-tang on the peri vascular immunoreactive nerve fiber of the basilar artery after experimentally induced subarachnoid hemorrhage(SAH). Sprague Dawley rats weighing between 350-400g were used. The SAH induced by injection of the fresh autologus heart blood(0.3-0.4ml) into the cisterna magna through the posterior atlanto-occipital membrane. Sample group was given a $3.3m{\ell}/kg/day$ of Yangkyuksanhoa-tang extracts for 2 days after SAH. The experimental animals divided into 48hrs after SAH. The changes of perivascular immunoreactive nerve fiber was examined by using indirect immunofluorescence method. The meshlike perivascular nerve fiber appeared in the basilar artery of normal rats. In basilar artery of SAH elicitated rat, the distribution of calcitonin gene-related peptide (CGRP)-immunoreactivity(IR) and vasoactive intestinal polypeptide(VIP)-IR of the perivascular nerve fiber were remarkably diminished, also dopamine beta hydroxylase(DBH)-IR, neuropeptide Y(NPY)-IR and serotonin-IR were diminished. In SAH elicitated rat with Yangkyuksanhoa-tang treatment, the CGRP-IR and VIP-IR degree were repaired as well as normal rat's, but DBH-IR, NPY-IR and serotonin-IR had no changes. These results provide the basic data to investigate the effect of Yangkyuksanhoa-tang on the vasospasm after SAH.

• Korean journal of applied entomology
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• v.32 no.4
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• pp.426-432
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• 1993

Thirteen isolates of Bacillus thunngiensls producing parasporal mclusions, obtained from 45 samples of dust and soil of sericultural tarms in Kyeong-ki province, were exammed for their toxicity against larvae of Lepidoptera, Dipwra and Coleoptera. Of these isolates, Bacillus f thuringiensis NT0423 was toxic to bath Lepidoptera, and Dipteran larvae. NT0423 showed that the $LC_{50}$ values against the Lepldaptora, Plutella macuhpennis and the Diptera, Culex pipiens were $1.30\times10^6$ CFU/ml, $2.88\times10^5$ CFU/ml, respectively. The tYPlcal bipyramldal crystals produced by NT0423 composed of protoxms of 130-140kDa encoded by one or more plasm mid-horne genes. Also, plasmid DNA analysis indicated Lhat this isolate has 9 plasmids which d differ with reported several B. thuringiensis strains.

• Korean journal of applied entomology
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• v.30 no.3
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• pp.180-186
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• 1991

This study was carried out to acquire some basic biochemical informations on the granulosis virus(GV) of Pieris rapae and Pieris brassicae. The capsule protein was composed of a single polypeptide with a molecular weight of 30,000 dalton for P. rapae GV and 31,000 dalton for P. brassicae GV. The major amino acids of capsule protein were glutamic acid, aspartic acid and lysine. When the capsule protein was partially digested with trypsin, chymotrypsin, papain or Staphylococcus aureus V8 protease, the digested products of the two viruses showed no difference in electrophoretic mobility. The patterns of the polypeptides of the two virus particle on SOS-polyacrylamide gel showed a little difference in high molecular weight region(over MW 100 kd).

• BMB Reports
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• v.37 no.4
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• pp.408-415
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• 2004

The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.241-247
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• 2016

Macrophage Migration Inhibitory Factor (MIF) are well-defined role as unique cytokine and critical mediator in acute and chronic inflammatory diseases, autoimmune diseases. In this study, we isolated and characterized a full-length of MIF cDNA from the abalone (Haliotis discus hannai). The full-length cDNA of abMIF was of 1264 bp, consisting of a 5'-terminal UTR of 143 bp, an open reading frame of 360 bp and a 3-terminal UTR of 761 bp. The abalone MIF cDNA encodes a 119-amino acid polypeptide with a calculated molecular mass of 13.4 kDa and isoelectric point of 9.07. Multiple alignments and phylogenetic analysis with the deduced abalone MIF protein and showed strong homology with disk abalone (Haliotis discusdiscus). The deduced amino acid sequence of abMIF exhibited homology with other reported MIFs, such as 80%, with that of other disk abalone H. discus discus MIF gene. Quantitative real-time PCR (qRT-PCR) analysis indicated that abMIF was highly expression observed in hapatopacreas, intestine, foot, and gonad of normal conditioned abalone. Even though AbMIF mRNA level in hemocytes was low under the normal condition, it was sharply up-regulated and reached the maximum at 6 h post-infection with Vibrio parahaemolyticus, and then decreased at 24 h post-infection. This result indicates that abMIF plays an important role in responding in the innate immune system.

• The Korean Journal of Zoology
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• v.34 no.4
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• pp.433-451
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• 1991

척추동물(7목 23종)의 췌장에서 insulin( B)세포, glucagon(A)세포, somatostatin( D)세포 및 pancreatic polypeptide( PP)세포 등을 면역세포화학법으로 동정하여 이들의 출현율, 분포양상 및 형태 등을 계통별로 비교하였다. 파충강의 거북목, 양서강의 유미목 및 어강의 악상대목 들을 제외한 모든 종에서 췌도의 형성을 관찰할 수 있었으며, 췌도를 구성하는 내분비세포의 크기에는 계통간의 차이가 있었다. B세포는 파충강의 것이 가장 크고, 양서강, 어강의 순이 었으며 A와 PP세포는 양서강, 파충강 및 어강의 순서였다. D세포는 양서강의 것이 가장 윤고, 다음이 어장이었으며, 파충강의 것이 가장 작았다. 이들 세포의 모양은 B세포의 경우 양서강과 어장에서는 원형, 난원형 및 방추형이었으며, 파충강에서는 원추형 및 단기형 등 다양한 모습이었다. A세포는 어강에서는 원헝, 난원형 및 방추형이 고르게 나타났고, 파충강과 양서 강에서는 원주형, 다각형 및 쐐기형이 나타났다. D세포는 모든 동물에서 원형, 난원형 및 방추형이 관찰되었고, 특히 파충강에서는 원추형및 쐐기형도 나타났다. PP세포는 주로 방추형 및 반원형이 대다수이였으며 간혹 원형 또는 다각형 등의 모습도 나타났다. 각 내분비세포의 출현율은 파충강 열 어강 들에서는 B, A, D 및 PP세포 순이었으나, 양서 강에서는 B, A, PP 및 D세포 순으로 나타났다. B와 PP세포는 양서강, 어강 및 파충강 순서로 출현하였고, A세포는 파충강, 어강 및 양서강의 순서이었으며 D세포는 어강, 파충강 및 양서강의 순서였다. 췌도 내에서의 세포의 분포 위치는 세포의 종류에 따라 차이를 보여 B세포의 경우 대다수 동물들에서 중앙부에 균등하게 분포하였으나 A, D및 PP세포는 주로 췌도 주변부에 분포하였고, 어강에서의 D세포는 췌도 중앙부에서도 관찰되었다. 일반적으로 파충강 및 양서 강에서는 외분비 선포조직에서초 내분비세포들이 출현하였으나, 어강에서는 내분비세포가 전혀 출현하지 않았다. 양서강 및 어강 들의 일부 수에서는 췌관상피에서도 드물게 나타났다.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.2
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• pp.223-229
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• 2011

Cyclophilins are originally identified as cytosolic binding protein of the immunosuppressive drug cyclosporine A. They have an activity of peptidyl prolyl cis/trans-isomerases (PPIase), which may play important roles in protein folding, trafficking, assembly and cell signaling. In this study, we report the cloning and characterization of a Bombyx mori cyclophilin A (bCypA) cDNA. The full-length cDNA of bCypA consist of 947 nucleotides with a polyadenylation signal sequence AATAAA and contain an open reading frame of 498 nucleotides encoding a polypeptide of 166 amino acids. The deduced amino acid sequence of bCypA shares a central peptidyl prolyl cis/trans-isomerase and a cyclosporin-A-binding domain with other cyclophilin sequences. Relative quantification real-time (RT) PCR analysis shows that mRNA transcripts of bCypA are detected in all the investigated tissues and highest expression level in the skin of 3-day-old 5 instar larva. Also, bCypA had PPIase activity on the proline-containing peptides. Accordingly, we suggest that bCypA is a new member of the cyclophilin A (CyPA) family and will be useful for quality control of bioactivity recombinant proteins with proline-containing peptides.