• International Journal of Industrial Entomology and Biomaterials
• /
• v.4 no.2
• /
• pp.101-107
• /
• 2002

We describe here the cDNA sequence and mRNA expression of a novel family of the antioxidant protein, peroxiredoxin, from the firefly, Pyracoetia ruin. The 555 bp cDNA sequence codes for a 185 amino acid protein with a calculated molecular mass of approximately 21 kDa. The deduced protein of P. rufa peroxiredoxin gene contains two conserved cysteine residues. Alignment of the deduced protein of P. rufa peroxiredoxin gene showed 71.1% protein sequenceidentity to known insect Drosophila melanogaster peroxiredoxin. Northern blot analysis revealed that the P. rufa peroxiredoxin is specifically expressed in the fat body of P. rufa larvae.

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.6
• /
• pp.871-880
• /
• 1999

The effects of dry roasting (110, 130, $150^{\circ}C$ for 15, 30, 45 min) on potential ruminant protein nutritional values in terms of: a), rumen bypass protein (BCP); b), rumen bypass starch (BST); c), fermented organic matter (FOM); d), true absorbed bypass protein (ABCP); e) microbial protein synthesized in the rumen based on available energy (E_MP); f), microbial protein synthesized in the rumen based on available nitrogen (N_MP); g), true protein supplied to the small intestine (TPSI); h), true absorbed rumen synthesized microbial protein (AMP); i), endogenous protein losses (ENDP); j), true digested protein in the small intestine (DVE); k), degraded protein balance (OEB) of whole lupin seeds (WLS) and faba beans (WFB) were evaluated by the new Dutch DV/OEB protein evaluation system. Dry roasting significantly increased BCP, BST, TPSI, ABCP, DVE (p<0.001) and decreased FOM, E_MP, AMP, N_MP and OEB (p<0.001) with increasing temperatures and times except that when temperature was at $110^{\circ}C$. The values of BCP, BST, TPSI, ABCP and DVE at $150^{\circ}C/45min$ for WLS and WFB were increased 2.2, 3.7; -, 2.0; 1.7, 1.7; 2.3, 3.7 and 1.7, 1.7 times and the values of FOM, E_MP, AMP, N_MP and OEB at $150^{\circ}C/45min$ for WLS and WFB were decreased by 15.3, 25.8; 18.1, 25.8; 18.7, 25.8; 54.6, 41.6 and 82.3% 54.7%, respectively, over the raw WLS and WFB. The results indicated that though dry roasting reduced microbial protein synthesis due to reducing FOM, TPSI didn't decrease but highly increased due to increasing BCP more than enough for compensation of the microbial protein decreasing. Therefore the net absorbable DVE in the small intestine was highly increased. The OEB values were significantly reduced for both WLS and WFB but not to the level of negative. It indicated that microbial protein synthesis might not be impaired due to the sufficient N supplied in the rumen, but the high positive OEB values in the most treatments except of $150^{\circ}C$ for 30 and 45 min of WLS (The OEB values: 54.8 and 26.0 g/kg DM) indicated that there were the large amounts of N loss in the rumen. It was concluded that dry roasting at high temperature was effective in shifting protein degradation from rumen to intestines and it increased the DVE values without reaching the negative OEB values. No optimal treatment was found in WLS due to the too high OEB values in all treatments. But dry roasting at $150^{\circ}C$ for 30 and 45 min might be optimal treatments for WLS due to the very lower OEB values.

• Journal of the Korean Magnetic Resonance Society
• /
• v.2 no.2
• /
• pp.131-140
• /
• 1998

The calcium-induced structural changes in the skeletal muscle regulatory protein troponin C (NTnC) involve a transition from a ‘closed’to an ‘open’structure with the concomitant exposure of a large hydrophobic interaction site for target proteins. Structural studies have served to define this conformational change and elucidate the mechanism of the linkage between calcium binding and the induced structural changes. There are now several structures of NTnC available from both NMR and X-ray crystallography. Comparison of the calcium bound structures reveals differences in the level of opening. We have considered the concept of a flexible open state of NTnC as a possible explanation for this apparent discrepancy. We also present simulations of the closed-to-open transition which are in agreement with the flexibility concept and with experimental energetics data.

• Korean Journal of Poultry Science
• /
• v.44 no.2
• /
• pp.67-73
• /
• 2017

The purpose of our study was to determine the effects of varying levels of energy, protein, and amino acids on the performances of laying hens. A total of 240 Hy-Line Brown laying hens at 36 weeks of age were used in this 4-week feeding trial. The hens were randomly allocated to five treatment diets, with eight replications of six hens in each replicate cage. The treatment diets were as follows: A- basal diet + 18% crude protein, metabolizable energy 2,800 kcal, total (methionine + cysteine) 0.65%; B- basal diet + 17% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; C- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; D- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.54%; and E- basal diet + 16% crude protein, metabolizable energy 2,680 kcal, total (methionine + cysteine) 0.54%. The study results revealed that the hen-day egg production of hens that were fed with low-energy diets (B, C, and D) was comparable with that of hens fed with high-energy diet A, whereas average daily feed intake in hens fed treatment diet D and E was significantly higher (P<0.05) than that in hens fed treatment diet A. Overall, the eggshell thickness was unaffected by any of the treatment diets. Egg weight was comparable among the treatment diets, except for treatment diet E. Haugh unit improved with decreasing levels of dietary energy, protein, and methionine + cysteine in the diet. We can summarize that laying hens fed with low dietary energy and low crude protein treatment diets B, C, and D had satisfactory performance compared with those fed with high-energy treatment diet A. This indicates that there is the potential to reduce feed costs by formulating diets with lower energy and low protein levels.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.3
• /
• pp.620-627
• /
• 2004

On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI- TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (enol). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic $\lambda$ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno 1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation.

• BMB Reports
• /
• v.38 no.5
• /
• pp.550-554
• /
• 2005

YKR049C is a mitochondrial protein in Saccharomyces cerevisiae that is conserved among yeast species, including Candida albicans. However, no biological function for YKR049C has been ascribed based on its primary sequence information. In the present study, NMR spectroscopy was used to determine the putative biological function of YKR049C based on its solution structure. YKR049C shows a well-defined thioredoxin fold with a unique insertion of helices between two $\beta$-strands. The central $\beta$-sheet divides the protein into two parts; a unique face and a conserved face. The 'unique face' is located between ${\beta}2$ and ${\beta}3$. Interestingly, the sequences most conserved among YKR049C families are found on this 'unique face', which incorporates L109 to E114. The side chains of these conserved residues interact with residues on the helical region with a stretch of hydrophobic surface. A putative active site composed by two short helices and a single Cys97 was also well observed. Our findings suggest that YKR049C is a redox protein with a thioredoxin fold containing a single active cysteine.

• Journal of Life Science
• /
• v.33 no.7
• /
• pp.531-537
• /
• 2023

Intracellular and axonal transport is mediated by microtubule-dependent motor proteins, such as kinesins and cytoplasmic dynein. Kinesin moves along the microtubule to the positive end of the microtubule, while dynein moves to the negative end of the microtubule. Kinesin-1 was first identified as a kinesin superfamily protein (KIF) that functions in the intracellular transport of various cargoes, including organelles, neurotransmitter receptors, and mRNA-protein complexes, through interactions between the carboxyl (C)-terminal domain and the cargo. It interacts with other cargoes, but the adapter/scaffold proteins that mediate between kinesin-1 and the cargo have yet to be fully identified. In this study, a yeast two-hybrid screen was used to identify adapter proteins that interact with the C-terminal region of KIF5A. We found an association between the C-terminal region of KIF5A and the cyclin-dependent kinase 2-associated protein 1 (CDK2AP1), originally identified in malignant hamster oral keratinocytes. CDK2AP1 bound to the C-terminal region of KIF5A and did not interact with KIF3A (the motor of kinesin-2), KIF5B, KIF5C, and kinesin light chain 1 (KLC1). The C-terminal region of CDK2AP1 is essential for its interaction with KIF5A. When co-expressed in HEK-293T cells, CDK2AP1 and kinesin-1 co-immunoprecipitated and co-localized in the cells. These results suggest that the KIF5A-CDK2AP1 interaction serves as an adapter protein connecting kinesin-1 and the cargo when kinesin-1 transports cargo in cells.

• Archives of Pharmacal Research
• /
• v.21 no.3
• /
• pp.315-319
• /
• 1998

Gastric smooth muscle of cats was used to investigate the involvement of protein kinase in vanadate-induced contraction. Vanadate caused a contraction of cat gastric smooth muscle in a dose-dependent manner. Vanadate-induced contraction was totally inhibited by 2 mM EGTA and 1.5 mM $LACI_3$ and significantly inhibited by $10\mu$M verapamil and $1\mu$M nifedipine, suggesting that vanadate-induced contraction is dependent on the extracellular $Ca^{2+}$ concentration, and the influx of extracellular $Ca^{2+}$ was mediated through voltage-dependent $Ca^{2+}$ channel. Both protein kinase C inhibitor and tyrosine kinase inhibitor significantly inhibited the vanadate-induced contraction and the combined inhibitory effect of two protein kinase inhibitors was greater than that of each one. But calmodulin antagonists did not have any influence on the vanadate-induced contraction. On the other hand, both forskolin ($1\mu$M) and sodium nitroprusside ($1\mu$M) significantly inhibited vanadate-induced contraction. Therefore, these results suggest that both protein kinase C and tyrosino kinase are involved in the vanadate-induced contraction which required the influx of extracellular $Ca^{2+}$ in cat gastric smooth muscle, and that the contractile mechanism of vanadate may be different from that of agonist binding to its specific receptor.

• IMMUNE NETWORK
• /
• v.1 no.3
• /
• pp.221-229
• /
• 2001

Background: Inactivation of tumor suppressor genes is believed to be important in the development of many human malignancies. Recently, several lines of evidence have indicated that the wild type p53 gene located at 17p13.3, may function as a tumor suppressor gene and that a mutant p53 gene could promote transformation by inactivating normal p53 function in a dominant negative fashion. These broad spectrum of p53 mutation in human cancers provide that mutant p53 and their protein may be potential targets of tumor diagnostic and therapeutic interventions. Method: Colony formation was performed to investigate growth suppressional ability. p53 expression pattern was examined by western blot and p53-mediated transactivation ability was assessed by CAT activity. SNU C2A cells were observed in apoptotic aspects induced by etoposide and $H_2O_2$ treatment, detecting sensitivity on agent, DNA fragmentation through agarose gel, chromatin condensation by fluorescence microscope, and cell cycle distribution by FACS. Result: 1) p53 mutant his179arg ($histidine{\rightarrow}arginine$) detected in SNU C2A cells lost transcriptional activity and growth suppression ability, showing dominant negative effect on its wild type p53. 2) Etoposide-treated SNU C2A cells induced apoptosis, exhibiting dramatic reduction of cell growth, DNA fragmentation, nuclear condensation formation of apoptotic body and increment of sub-G1 cell fraction. 3) Etoposide and $H_2O_2$-treated SNU C2A cells have no high increase of p53 expression and overexpressed p53 protein changed localization, from cytoplasm to nucleus. Also, p53-mediated transcriptional activity was increased by agents-treatment. Conclusion: SNU C2A cells coexpress wild-type and mutant p53 protein induced apoptosis in the condition on DNA damage, through localizational shift from cytoplasm to nucleus of p53 protein rather than the induction of p53 protein. SNU C2A cells derived mutant p53 his179arg abrogated both the growth supression ability and transactivational activity, showing inhibition effect on transcriptional activity of wild type p53, but did not repress the activity of wild type p53 in SNU C2A cells owing to dominant activity of wild type. These cell condition may provide new gene therapeutic implications leading effective antiproliferation of cell when mutant and wild-type p53 protein were co-expressed in cell.

• Journal of Nutrition and Health
• /
• v.6 no.2
• /
• pp.35-43
• /
• 1973

This study is concerned with the nutritional status of the packed lunch boxes which are brought by the junior high school students in Capital City of Seoul. Four hundred and eights students of the 8 school districts in Seoul had been randomly selected as subjects. The contained nutrients in the packed lunch were analysed by the Food Composition Table. To observe the influence of home economical status and mother's educational level on the nutrient concents of packed lunches, the chosen subjects were classified into three large groupings, which are upper, middle and low classes respectively. In addition, comparison between the Recommended Daily Allowanced for Korean people-13 to 15 age group-and the contained nutrients in the lunch boxed had been conducted. T-test was applied to clarify the significance of the differences between each group both economical and educational level. 1. The averaged rations between the Recommended Daily Allowances and the contained nutrients in the lunches stand: Calorie 59.7% (566 Cal.) protein 53.1% (18g), animal protein 48.6% (5g), fat 39.8% (5g), calcium 371.% (0.1g), ferret 66.4% (2.9g) Vitamin A 642 1.U. (31.3%), Vitamin $B_1$ 70.2% (0.3mg), Vitamin $B_2$ 41.4% (0.2g), Niacin 77.0%(4mg), Vitamin C 51.9% (13mg). All of the nctrients fall far behind the Recommended Daily, Allowances for 13 to 15 age group. 2. Home economical status brings influence on the kinds of foods which could been. Protein, animal protein, fat, Vitamin $B_2$, Vitamin C showed significance of diffierences between the upper and middle classes. Between the middle and low classes, Protein, animal protein, fat, calcium and Vitamin C showed significance of difference. And finally, between the upper and low classes, protein, fat, calcium, ferret, Vitamin $B_2$ and Vitamin C showed a great significance. 3. Regardless to the living conditions of the subject students, all the nutrients of the lunches packed by the mothers in the entire educational levels did not reach the Recommended Daily Allowances. Protein, animal protein, fat, ferret, Vitamin A, and Niacin showed the significance of the differences between the upper and middle classes. On the other hand, calorie, animal protein, fat, Vitamin A and Vitamin C showed the significance between the middle and low classes. Between the upper and the low classes, protein, animal protein, fat, and ferret showed significance. 4. The mairdish-ice of the lunch boxes supplied calorie, protein, Vitamin $B_1$, Vitamin $B_2$ and Niacin which stand at 83.8%, 56.1%, 52.5%, and 54.8% respectivly when compared with the whole nutritional contents. 5. The side-dishes of the packed lunch lack in variety of cooking methods. One interesting fact is that entire subject students are very favorable to the food cooked with every kind of grains.