• Fisheries and Aquatic Sciences
• /
• 제16권4호
• /
• pp.311-318
• /
• 2013

The gene encoding an esterase from Photobacterium sp. MA1-3 was cloned in Escherichia coli using the shotgun method. The amino acid sequence deduced from the nucleotide sequence (948 bp) corresponded to a protein of 315 amino acid residues with a molecular weight of 35 kDa and a pI of 6.06. The deduced protein showed 74% and 68% amino acid sequence identities with the putative esterases from Photobacterium profundum SS9 and Photobacterium damselae, respectively. Absence of a signal peptide indicated that it was a cell-bound protein. Sequence analysis showed that the protein contained the signature G-X-S-X-G included in most serine-esterases and lipases. The MA1-3 esterase was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme was a serine-esterase and was active against $C_2$, $C_4$, $C_8$ and $C_{10}$ p-nitrophenyl esters. The optimum pH and temperature for enzyme activity were pH 8.0 and $30^{\circ}C$, respectively. Relative activity remained up to 45% even at $5^{\circ}C$ with an activation energy of 7.69 kcal/mol, which indicated that it was a cold-adapted enzyme. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, $Zn^{2+}$, and $Hg^{2+}$ ions.

• Molecular & Cellular Toxicology
• /
• 제1권4호
• /
• pp.275-280
• /
• 2005

Carbon tetrachloride ($CCl_4$) is well known hepatotoxicant. Its overdose cause severe centrilobular hepatic necrosis in human and experimental animals. We administered $CCl_{4}$ at low (0.2 mL/kg p.o.) and high (2 mL/kg p.o.) doses to mice. Mice were sacrificed at 24 h after administration. We evaluated liver toxicity by serum AST and ALT level and by microscopic observation. Using cDNA chip, we conducted gene expression analysis in liver. Mean serum activities of the hepatocellular leakage enzymes, ALT and AST, were significantly increased compare to control, respectively, in the low and high dose groups. H&E evaluation of stained liver sections revealed $CCl_{4}-related$ histopathological findings in mice. Moderate centrilobular hepatocellular necrosis was present in all $CCl_{4}$ treated mice. We found that gene expression pattern was very similar between low and high dose group. However, some stress related genes were differently expressed. These results could be a molecular signature for the degree of liver injury. Our data suggest that the degree of severity could be figure out by gene expression profiling.

• Biomolecules & Therapeutics
• /
• 제25권3호
• /
• pp.239-248
• /
• 2017

Desensitization and acute tolerance are terms used to describe the attenuation of receptor responsiveness by prolonged or intermittent exposure to an agonist. Unlike desensitization of G protein-coupled receptors (GPCRs), which is commonly explained by steric hindrance caused by the ${\beta}$-arrestins that are translocated to the activated receptors, molecular mechanisms involved in the acute tolerance of GPCRs remain unclear. Our studies with several GPCRs and related mutants showed that the acute tolerance of GPCRs could occur independently of agonist-induced ${\beta}$-arrestin translocation. A series of co-immunoprecipitation experiments revealed a correlation between receptor tolerance and interactions among receptors, ${\beta}$-arrestin2, and $G{\beta}{\gamma}$. $G{\beta}{\gamma}$ displayed a stable interaction with receptors and ${\beta}$-arrestin2 in cells expressing GPCRs that were prone to undergo tolerance compared to the GPCRs that were resistant to acute tolerance. Strengthening the interaction between $G{\beta}{\gamma}$ and ${\beta}$-arrestin rendered the GPCRs to acquire the tendency of acute tolerance. Overall, stable interaction between the receptor and $G{\beta}{\gamma}$ complex is required for the formation of a complex with ${\beta}$-arrestin, and determines the potential of a particular GPCR to undergo acute tolerance. Rather than turning off the signal, ${\beta}$-arrestins seem to contribute on continuous signaling when they are in the context of complex with receptor and $G{\beta}{\gamma}$.

• BMB Reports
• /
• 제45권3호
• /
• pp.133-140
• /
• 2012

Cancers claim millions of lives each year. Early detection that can enable a higher chance of cure is of paramount importance to cancer patients. However, diagnostic tools for many forms of tumors have been lacking. Over the last few years, studies of chimeric RNAs as biomarkers have emerged. Numerous reports using bioinformatics and screening methodologies have described more than 30,000 expressed sequence tags (EST) or cDNA sequences as putative chimeric RNAs. While cancer cells have been well known to contain fusion genes derived from chromosomal translocations, rearrangements or deletions, recent studies suggest that trans-splicing in cells may be another source of chimeric RNA production. Unlike cis-splicing, trans-splicing takes place between two pre-mRNA molecules, which are in most cases derived from two different genes, generating a chimeric non-co-linear RNA. It is possible that trans-splicing occurs in normal cells at high frequencies but the resulting chimeric RNAs exist only at low levels. However the levels of certain RNA chimeras may be elevated in cancers, leading to the formation of fusion genes. In light of the fact that chimeric RNAs have been shown to be overrepresented in various tumors, studies of the mechanisms that produce chimeric RNAs and identification of signature RNA chimeras as biomarkers present an opportunity for the development of diagnoses for early tumor detection.

• 한국대기환경학회지
• /
• 제31권5호
• /
• pp.482-492
• /
• 2015

Ground-based measurements were conducted from January 6 to 12 of 2015 for understanding characteristics of nitrogen containing carbonaceous aerosols as 16 amino acids at the Mokpo National University, Korea. The detailed amino acid components such as Cystine ($(SCH_2CH(NH_2)CO_2H)_2$) and Methionine ($C_5H_{11}NO_2S$) and their sources were analyzed by High-Performance Liquid Chromatography with Fluorescence Detection (HPLC-FLD) for behavior of secondary products in particulate matter. In addition, organic carbon (OC) and elemental carbon (EC) based on the carbonaceous thermal distribution (CTD), which provides detailed carbon signature characteristics relative to analytical temperature, and water soluble organic carbon (WSOC) by total organic carbon (TOC) analyzer were used to understand the carbon compound behaviors. The backward trajectories were discussed for originations of carbonaceous aerosols as well. Different airmasses were classified with the amino acids and OC thermal signatures. The results can provide to understand the aging process influenced by the long-range transport from East Sea area.

• 한국어병학회지
• /
• 제28권2호
• /
• pp.99-107
• /
• 2015

Despite its economic importance as an aquaculture species, the molecular and genetic information regarding physiologically important elements in rock bream (Oplegnathus fasciatus) is not completely understood. Rab proteins play a vital role in cellular mechanisms and immunity as one of the key regulators of membrane trafficking. In this investigation, a Rab gene, named as RbRab-5C-like, was identified from Oplegnathus fasciatus. RbRab-5C-like protein exhibited high homology with Rab proteins of other species and possessed signature characteristics of Rab proteins with four conserved cysteine residues. Phylogenetic analysis showed that RbRab-5C-like clustered with other fish counterparts. The RbRab-5C-like genomic sequence possesses six exons and five introns. Transcriptional analysis revealed that RbRab-5C-like was ubiquitously expressed in all examined tissues with the highest expression occurring in the liver. While the structural and homologic characteristics of RbRab-5C-like suggest a strong conservation of this element in different species, its mRNA distribution implies a wide range of biological significance in rock bream.

• 한국대기환경학회지
• /
• 제29권6호
• /
• pp.722-733
• /
• 2013

Ground-based measurements were conducted from August 25 to September 8 of 2011 for understanding characteristics of carbonaceous aerosols measured at Gosan. Chemical components and their sources were discussed by analysis of organic compounds with identification of primary and secondary products in particulate matter. Thus, organic carbon (OC) and elemental carbon (EC) based on the carbonaceous thermal distribution (CTD), which provides detailed carbon signature characteristics relative to analytical temperature, was used to improve the carbon fractionation of the analytical method. In addition, organic compounds by gas chromatography technique with the backward trajectories were discussed for characteristics of carbonaceous aerosols. Different air-masses were classified related to the OC thermal signatures and the organic molecular markers such as aromatic acids and PAHs. We concluded that the aging process was influenced by the long-range transport from East Sea area.

• BMB Reports
• /
• 제39권5호
• /
• pp.578-585
• /
• 2006

The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMP-binding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.

• The Plant Pathology Journal
• /
• 제15권3호
• /
• pp.131-136
• /
• 1999

In fungi known as ascomycetes, ability to mate is controlled by a single mating type (MAT) locus with two dissimilar sequences called idiomorphs carrying genes encoding transcription factors that are unrelated to each other. Fungi requiring strains with different MAT genes to complete the sexual process are heterothallic (self-sterile); species in which as single strain is able to undergo sexual reproduction are homothallic (self-fertile). Previous analysis of sequences from several heterothallic and homothallic species of the ascomycete genus Cochliobolus showed that homothallics evolve from heterothallics and that each known Cochliobolus homothallic species arose independently, from a different heterothallic ancestral species. Here we report detailed comparative analyses of MAT sequences ad their flanking regions, and show that: (1) The level of MAT gene similarity is not correlated with reproductive life style; (2) MAT proteins from all Cochliobolus species are conserved within the transcription factor signature sequences; they are not conserved in the carboxy terminal half of MAT-1, or third of MAT-2, except in those from very closely related species; (3) A gene (ORF1) of unknown function, consistently found on the MAT flank, is more conserved than are the MAT genes themselves; (4) The intergenic sequences diverge sharply among species.

• Molecules and Cells
• /
• 제39권2호
• /
• pp.77-86
• /
• 2016

Cancer is a heterogeneous disease caused by diverse genomic alterations in oncogenes and tumor suppressor genes. Despite recent advances in high-throughput sequencing technologies and development of targeted therapies, novel cancer drug development is limited due to the high attrition rate from clinical studies. Patient-derived xenografts (PDX), which are established by the transfer of patient tumors into immunodeficient mice, serve as a platform for co-clinical trials by enabling the integration of clinical data, genomic profiles, and drug responsiveness data to determine precisely targeted therapies. PDX models retain many of the key characteristics of patients' tumors including histology, genomic signature, cellular heterogeneity, and drug responsiveness. These models can also be applied to the development of biomarkers for drug responsiveness and personalized drug selection. This review summarizes our current knowledge of this field, including methodologic aspects, applications in drug development, challenges and limitations, and utilization for precision cancer medicine.