• Title/Summary/Keyword: X protein

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Ribosomal Protein S4 Genes in Macaca fuscata: Sequence, Evolution, and Phylogeny

  • Kim, Heui-Soo
    • Journal of Life Science
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    • v.11 no.1
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    • pp.34-38
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    • 2001
  • The cDNA encoding ribosomal protein S4(RPS 4) from an ovary cDNA library of the Japanese monkey (Macaca fuscata) was cloned and sequenced. The RPS4X gene from monkey X chromosome encodes a deduced protein of 263 amino acids and share 99.1% cDNA sequence similarity and 100% amino acid sequence identify with the human RPS4X. Rate of synonymous substitution was higher in RPS4Y than in RPS4X in comparison to the monkey and human. The ratio of synonymous and nonsynonymous substitutions per site indicated that directional selection has nor occurred in RPS4 genes. Phylogenetic analysis using the neighbor-joining method revealed that X and Y-linked RPS4 genes have evolved independently.

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Assessment of the Specificity of A Hybridization of Surfactant Protein A by Addition of Non-specific Rat Spleen RNA (Surfactant Protein A mRNA을 이용한 유전자 재결합 반응에서 비특이성 RNA의 첨가에 의한 특이성 검정)

  • Kim, Byeong Cheol;Kim, Mi Ok;Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.4
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    • pp.393-404
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    • 2004
  • Background : Nucleic acid hybridization has become an essential technique in the development of our understanding of gene structure and function. The quantitative analysis of hybridization has been used in the measurement of genome complexity and gene copy number. The filter hybridization assay is rapid, sensitive and can be used to measure RNAs complementary to any cloned DNA sequence. Methods : The authors assessed the accuracy, linearity, correlation coefficient and specificity of the hybridization depending on the added dose(0, 1, 5, and $10{\mu}g$) of non-specific rat spleen RNA to hybridization of surfactant protein A mRNA. Filter hybridization assays were used to obtain the equation of standard curve and thereby to quantitate the mRNA quantitation. Results : 1. Standard curve equation of filter hybridization assay between counts per minute (X) and spleen RNA input (Y) was Y=0.13X-19.35. Correlation coefficient was 0.98. 2. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) was Y=0.00066X-0.046. Correlation coefficient was 0.99. 3. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $1{\mu}g$ spleen RNA was Y=0.00056X-0.051. Correlation coefficient was 0.99. 4. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $5{\mu}g$ spleen RNA was Y=0.00065X-0.088. Correlation coefficient was 0.99. 5. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $10{\mu}g$ spleen RNA was Y=0.00051X-0.10. Correlation coefficient was 0.99. Conclusions : Comparison of cpm/filter in a linear range allowed accurate and reproducible estimation of surfactant protein A mRNA copy number irrespective of the addition dosage of non-specific rat spleen RNA over the range $0-10{\mu}g$.

Docking Studies on Formylchromone Derivatives as Protein Tyrosine Phosphatase 1B (PTP1B) Inhibitors

  • Kim, Chan-Kyung;Lee, Kyung-A;Zhang, Hui;Cho, Hyeong-Jin;Lee, Bon-Su
    • Bulletin of the Korean Chemical Society
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    • v.28 no.7
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    • pp.1141-1150
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    • 2007
  • Molecular modeling study has been performed to assist in the design of PTP1B inhibitors using FlexX. FlexX dockings with 19 test ligands, whose structures have been determined by X-ray crystallography, were successful in reproducing the experimental conformations within the protein. An increase in biological activity is observed as hydrophobic character of formylchromone derivatives increases. Most ligands bind to the activesite regions of the protein successfully in two different score runs. The Drug score run gave better results than the FlexX score run based on the score, rank, binding modes and bond distance of docked structures. Consensus values from the CScore scoring function are between 3 and 5, suggesting that the scoring scheme is reliable. All formylchromone inhibitors considered in this work show unidirectional binding modes in the active site pocket, which is contrary to the bidirectional X-ray results by Malamas et al. and amino acid residues responsible for such orientation are identified to help further development of the inhibitors.

Analysis of Toxicity in Escherichia coli from the Expression of Human Purinergic Receptor $P2X_4$ (인간 퓨린수용체 $P2X_4$를 발현시킬 때 나타나는 대장균 독성의 원인분석)

  • Yu, Yon-Joo;Jung, Yun-A;Lim, Dong-Bin
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.7-13
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    • 2011
  • In general, expression of membrane protein in Escherichia coli is very toxic to the host organism, but the mechanism for the toxicity is not clear yet. Expression of human purinergic receptor $P2X_4$ was found to be extremely toxic to the host E. coli. We examined this toxicity by isolation and analysis of less toxic mutant proteins. We could isolate 30 less toxic mutants of $P2X_4$ after hydroxylamine mutagenesis. Western blot showed that all of them produced proteins smaller than the wild type $P2X_4$. DNA sequencing of two largest mutant proteins showed that they were lost its second transmembrane domain. Localization analysis of these mutant proteins showed that they are not in cytoplasmic membrane, but in inclusion bodies. These data showed that inactive truncated $P2X_4$ is not toxic to E. coli and membrane integration and functionality of $P2X_4$ may be needed to show host toxicity.

Isolation of $\beta$-Lactamase Inhibitory Protein from Streptomyces exfoliatus SMF19 and Cloning of the Corresponding Gene

  • PARK, HYEON-UNG;KYE JOON LEE
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.369-374
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    • 1996
  • The ${\beta}$-lactamase inhibitory protein (BLIP) produced by Streptomyces exfoliatus SMF19 was purified(33 kDa) and the N-terminal amino acid sequence was determined as NH2-ATSVVAWGGNND. Genomic DNA library of S. exfoliatus SMF19 was constructed in pWE15 and recombinants harbouring the corresponding gene were selected by colony hybridization to the mixture of 36-mer oligonucleotide designed from the N-terminal amino acid sequence. The corresponding gene (bliX) was isolated on a 4-kb ApaI fragment of S. exfoliatus SMF19 chromosomal DNA and then sequenced. The bliX consisting of 1, 119bp encoded a mature protein with a deduced amino acid sequence of 342 residues and also encoded a 40-amino-acid signal sequence. No significant sequence similarity to bliX was found by pairwise comparison using various protein and nucleotide sequences.

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THE UTILIZATION OF DIETARY PROTEIN BY YOUNG RAINBOW TROUT (무지개송어의 사료단백질 이용에 관한 연구)

  • KIM Yong Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.7 no.4
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    • pp.209-214
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    • 1974
  • The utilization dietary protein in young rainbow trout was investigated when feeded with the diets of controlled levels of casein and lipids for 12 days. The composition of the diet is shown in Table 1. Body weight gain was in proportion to the protein content in diet up to $40\%$, ana the maximum was obtained with $40.4\%$ of crude protein (Fig. 1, Table 2). On the other hand, the accumulated protein in body also showed almost maximum value around 40n of protein level in the diet and: the change of accumulated protein showed a little with more-protein level(Fig. 3, Table 9). The protein accumulation rate (protein accumulated/protein consumed) showed its highest value at about $40\%$ of protein level, while the protein utilization value (protein accumulation rate x protein content of diet) attained its highest value at $54.8\%$ of protein level (Table 3). With the above results it is observed that the requirement of dietary protein for young rainbow trout is about $40\%$ of protein in the diet when casein is used as the sole protein source. In protein efficiency ratio (PER) the lower protein level in the diet, the higher PER yields and the more the quantity of protein increases, the more PER decreases and its relation could be figured out as an equation of y=4.91-0.034x (Fig.4), Nose measured PER utilizing the diet which result of this, it is reported that PER rate of casein dropped within the extent of $25\%$ protein in diet. The reason why such a different PER rate appeared at the low protein level is revealed as the carbohydrate is low but the lipid high in capacity of utililzing nutrients for rainbow trout. The relation between the protein content of diet and the conversion factor, feed efficiency were determined and the results are shown in Fig. 2.

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Urine Specific Gravity as a Useful Tool for Screening Proteinuria in Children (소아 단백뇨 검사에 있어서 요비중의 유용성)

  • Kim, Jong-Hwa;Yoo, Kee-Hwan;Hong, Young-Sook;Lee, Joo-Won;Kim, Soon-Kyum
    • Childhood Kidney Diseases
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    • v.4 no.1
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    • pp.1-5
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    • 2000
  • Purpose: The plcr of spot urine has been uised to predict the timed urine protein excretion. Although this method reduces errors caused by variations in urine volume, it is relatively thconvenient and expensive. Recently, a more rapid and less expensive screening method with specific gravity(SG) has been reported, and we have examined whether estimated-creatinine(Cr-est) with urine 5G could be used in place of urine creatinine to predict 24-hour collected urine protein excretion in children. Methods: We had retrospectively analyzed protein, creatinine and urine SG in randomized spot urine samples of 147 patients from March 1998 till June 1998 in Korea university Guro hospital and compared the urinary protein creatinine ratio(P/Cr) with the protein estimated-creatinine ratio(P/Cr-est). We compared the correlation of urinary creatinine vs-urine 5G with the timed urine pretein excretion. Results: 1) urine SG accurately estimated urine creatinine concentration (r=0.407, P<0.001, Cr=SG x 4485.82-4482.87). 2) P/Cr correlated with urine protein excretion measured in a 24-hour urine collection (r=0.771, P<0.001, 24-hour collected urine protein : 0.338 x (P/Cr) 4+667.885). 3) P/Cr-est correlated with a 24-hour collected urine protein (r=0.723, P<0.001, 24-hour collected urine protein =0.354 x (P/Cr-est)+726.044), Conclusions: These results suggest that P/Cr-est with urine SG could be useful method for screening proteinuria in children.

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STUDIES ON BIOCHEMICAL POLYMORPHISM OF MILK PROTEIN AS GENETIC MARKERS IN PIGS

  • Chung, E.R.;Han, S.K.;Shin, Y.C.;Chung, H.Y.;Kim, J.E.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.2
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    • pp.285-294
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    • 1992
  • Biochemical polymorphisms of sow's milk proteins, $\beta$-casein ($\beta$-CN), $\beta$-lactoglobulin ($\beta$-LG), post-lactoglobulin (post-LG), $\alpha$-lactalbumin ($\alpha$-LA) and X-protein, as genetic markers for major pig breeds (Landrace, Yorkshire, Duroc, Hampshire and cross bred) in Korea were determined by starch gel electrophoresis. Phenotype and gene frequencies at all marker loci were estimated and genetic differences among breed populations were analyzed. Three $\beta$-CN phenotypes (AA, AB and BB) controlled by two codominant alleles (${\beta}-CN^A$ and ${\beta}-CN^B$), four $\beta$-LG phenotypes (AA, AC, $AC^{\pm}$ and CC) controlled by two codominant alleles (${\beta}-LG^A$ and ${\beta}-LG^C$) and ten X-protein phenotypes (AA, BB, CC, DD, AB, AC, AD, BC, BD and CD) controlled by four codominant alleles ($X^A,\;X^B,\;X^C\;and\;X^D$) were identified. In addition, a genetically controlled polymorphism of post-LG was found for the first time in sow's milk protein. Three different phenotypes (AA, AB and BB) were designated $post-LG^A$ and $post-LG^B$. Of the five marker loci examined, $\alpha$-LA locus was observed to lack any individual variation in all breeds studied. All populations were in Hardy-Weinberg equilibrium for all loci. There were marked breed differences for phenotype and gene frequencies in the post-LG and X-protein marker loci. However, there were little differences between breeds in the gene frequencies at the $\beta$-CN and $\beta$-LG marker loci.

Altered Translational Control of Fragile X Mental Retardation Protein on Myelin Proteins in Neuropsychiatric Disorders

  • Jeon, Se Jin;Ryu, Jong Hoon;Bahn, Geon Ho
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.231-238
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    • 2017
  • Myelin is a specialized structure of the nervous system that both enhances electrical conductance and insulates neurons from external risk factors. In the central nervous system, polarized oligodendrocytes form myelin by wrapping processes in a spiral pattern around neuronal axons through myelin-related gene regulation. Since these events occur at a distance from the cell body, post-transcriptional control of gene expression has strategic advantage to fine-tune the overall regulation of protein contents in situ. Therefore, many research interests have been focused to identify RNA binding proteins and their regulatory mechanism in myelinating compartments. Fragile X mental retardation protein (FMRP) is one such RNA binding protein, regulating its target expression by translational control. Although the majority of works on FMRP have been performed in neurons, it is also found in the developing or mature glial cells including oligodendrocytes, where its function is not well understood. Here, we will review evidences suggesting abnormal translational regulation of myelin proteins with accompanying white matter problem and neurological deficits in fragile X syndrome, which can have wider mechanistic and pathological implication in many other neurological and psychiatric disorders.

The hepatitis B virus X protein induced fibrosis in Huh7 cells (간암세포주 Huh7에서 Hepatitis B virus X protein에 의한 간섬유화)

  • Son, Moa;Park, Sanggyu;Cho, Moonjae
    • Journal of Applied Biological Chemistry
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    • v.59 no.1
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    • pp.25-29
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    • 2016
  • Hepatitis B virus infection can cause hepatic fibrosis leading to cirrhosis and hepatocellular carcinoma. However the mechanism remains poorly understood. In this study, we found that Hepatitis B virus X-protein (HBx) increases vimentin, fibronectin, slug, snail and NOX4 expression. Because NOX4-mediated reactive oxygen species can increase slug and snail, which can induce fibrosis, HBx may be a key regulator of hepatic fibrosis development via NOX4 induction.