• Development and Reproduction
• /
• v.26 no.1
• /
• pp.1-12
• /
• 2022

This study aimed to investigate the signal transduction of phosphorylation sites at the carboxyl (C)-terminal region of equine luteinizing hormone/chorionic gonadotropin receptor (eLH/CGR). The eLH/CGR has a large extracellular domain of glycoprotein hormone receptors within the G protein-coupled receptors. We constructed a mutant (eLH/CGR-t656) of eLH/CGR, in which the C-terminal cytoplasmic tail was truncated at the Phe656 residue, through polymerase chain reaction. The eLH/CGR-t656 removed 14 potential phosphorylation sites in the intracellular C-terminal region. The plasmids were transfected into Chinese hamster ovary (CHO)-K1 and PathHunter Parental cells expressing β-arrestin, and agonist-induced cAMP responsiveness was analyzed. In CHO-K1 cells, those expressing eLH/CGR-t656 were lower than those expressing eLH/CGR wild-type (eLH/CGR-wt). The EC₅₀ of the eLH/CGR-t656 mutant was approximately 72.2% of the expression observed in eLH/CGR-wt. The maximal response in eLH/CGR-t656 also decreased to approximately 43% of that observed in eLH/CGR-wt. However, in PathHunter Parental cells, cAMP activity and maximal response of the eLH/CGR-t656 mutant were approximately 173.5% and 100.8%, respectively, of that of eLH/CGR-wt. These results provide evidence that the signal transduction of C-terminal phosphorylation in eLH/CGR plays a pivotal role in CHO-K1 cells. The cAMP level was recovered in PathHunter Parental cells expressing β-arrestin. We suggest that the signal transduction of the C-terminal region phosphorylation sites is remarkably different depending on the cells expressing β-arrestin in CHO-K1 cells.

• Korean Journal of Ichthyology
• /
• v.13 no.1
• /
• pp.24-27
• /
• 2001

The barbel structure of the spined loach Iksookimia longicorpa were investigated histologically. Their barbels consist of epidermis, dermis, and a central rod. The epidermis contains mucous cells, terminal buds, granular cells, and epidermal cells. The mucous cells are thin rims of basophilic cytoplasm and are located at the surface of the epidermis. The terminal bud is basophilic and is situated at the distal portion of the epidermis. The dermis consists of loose connective tissue containing blood vessels, pigment cells, and nerve cells. The central region of cartilage is the innermost region and is enclosed within muscle layers.

• Korean Journal of Microbiology
• /
• v.42 no.3
• /
• pp.165-171
• /
• 2006

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.10
• /
• pp.1756-1762
• /
• 2007

Envelope proteins of virus contain a segment of hydrophobic amino acids, called as fusion peptide, which triggers membrane fusion by insertion into membrane and perturbation of lipid bilayer structure. Potential fusion peptide sequences have been identified in the middle of L or M proteins or at the N-terminus of S protein in the envelope of human hepatitis B virus (HBV). Two 16-mer peptides representing the N-terminal fusion peptide of the S protein and the internal fusion peptide in L protein were synthesized, and their membrane disrupting activities were characterized. The internal fusion peptide in L protein showed higher activity of liposome leakage and hemolysis of human red blood cells than the N-terminal fusion peptide of S protein. Also, the membrane disrupting activity of the extracellular domain of L protein significantly increased when the internal fusion peptide region was exposed to N-terminus by the treatment of V8 protease. These results indicate that the internal fusion peptide region of L protein could activate membrane fusion when it is exposed by proteolysis.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.586-589
• /
• 2003

Previously we reported the cloning and sequence analysis of a CFTase gene from Bacillus polymyxa. CFTase was divided into five distinct regions. In order to understand a role of the N-terminal repeat region on the function of CFTase from Bacillus polymyxa MGL21, deletion mutantCFTase ${\Delta}N$ was prepered. Recombinant protein was overproduced in E. coli as inclusion body, solubilized in bufer containing 8M urea, and refolded in the phosphate buffer. The molecular weight of the purified wild type CFTase and CFTase ${\Delta}N$ were 148kDa , 108kDa, respectively.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.143.2-143
• /
• 2003

A rapid and sensitive assay for the specific detection of plant viroids using reverse transcription-polymerase chain reaction(RT-PCR) has been developed already. The nested RT-PCR assay cloud be applied for the detection of apple scar skin viroid(ASSVd) from young leaves and other tissues. ASSVd has central conserved region(CCR), terminal left(T$\sub$L/) and terminal right(T$\sub$R/) domain. Primers were designed from these regions. Primer sets were successfully applicable for the amplification of full length or partial region of ASSVd by nested RT-PCR. Nested RT-PCR assay was more sensitive and accurate method to detect ASSVd from young trees during the early time of apple cultivation.

• Journal of Applied Biological Chemistry
• /
• v.53 no.2
• /
• pp.112-115
• /
• 2010

To analyze the surface properties of bacteriophytochrome (BphP), five (2B8, 2C11, 3B2, 3D2, 3H7) anti-BphP monoclonal antibodies were produced by using full-length of BphP of Deinococcus radiodurnas. 2B8 and 2C11 preferentially recognized the epitopes at N-terminal region of BphP, whereas 3B2, 3D2 and 3H7 showed preferential affinities to the epitopes of C-terminal region of BphP.

• The Journal of Korean Society of Virology
• /
• v.30 no.3
• /
• pp.211-217
• /
• 2000

Nef is a lentiviral protein involved in pathogenesis of AIDS, but its molecular mechanism of action remains incompletely understood. Here we report the isolation of the interacting protein with the HIV-1 Nef, using the yeast two hybrid system for expression cloning. One of the positive colonies was selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed that this interacting protein is Human Ikaros/LyF-1. This protein interacted with the C-terminal region of Nef specifically in yeast system, not with the N-terminal region. This interaction was also confirmed by in vitro binding assay.

• Genomics & Informatics
• /
• v.6 no.3
• /
• pp.130-135
• /
• 2008

The haplotypes of a 200 kb region in the human chromosome X terminal band (q28) were analyzed using the International HapMap Project Phasell data, which had been collected for three analysis panels (YRI, CEU, and CHB＋JPT). When multiple linkage disequilibrium blocks were encountered for a panel, the neighboring haplotypes that had crossover rate of 5% or more in the panel were combined to generate 'haploid' configurations. This resulted in 8, 7, and 5 'haploid' configurations for the panels of YRI, CEU, and CHB＋JPT, respectively. The multiple sequence alignment of these 'haploids' was used for the calculation of allele-sharing distances and the subsequent principal coordinate analysis. Two 'haploids' in CEU and CHB＋JPT were hypothesized as 'parental' in light of the observations that the successive recombinants of these haploids can model two other haploids in CEU and CHB＋JPT, and that their configurations were consistent with those in YRI. This study demonstrates the utility of haplotype phylogeny in understanding population evolution.

• Journal of the Korean Institute of Telematics and Electronics D
• /
• v.35D no.12
• /
• pp.53-58
• /
• 1998

A buried channel poly-Si TFT (BCTFT) for application of high performance integrated circuits has been proposed and fabricated. BCTFT has unique features, such as the moderately-doped buried channel and counter-doped body region for conductivity modulation, and the fourth terminal entitled back bias for preventing kink effect. The n-type and p-type BCTFT exhibits superior performance to conventional poly-Si TFT in ON-current and field effect mobility due to moderate doping at the buried channel. The OFF-state leakage current is not increased because the carrier drift is suppressed by the p-n junction depletion between the moderately-doped buried channel and the counter-doped body region.