• BMB Reports
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• v.38 no.3
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• pp.275-280
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• 2005

For most of proteins to be active, they need well-defined three-dimensional structures alone or in complex. Folding is a process through which newly synthesized proteins get to the native state. Protein folding inside cells is assisted by various chaperones and folding factors, and misfolded proteins are eliminated by the ubiquitin-proteasome degradation system to ensure high fidelity of protein expression. Under certain circumstances, misfolded proteins escape the degradation process, yielding to deposit of protein aggregates such as loop-sheet polymer and amyloid fibril. Diseases characterized by insoluble deposits of proteins have been recognized for long time and are grouped as conformational diseases. Study of protein folding mechanism is required for better understanding of the molecular pathway of such conformational diseases.

• Journal of Life Science
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• v.19 no.12
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• pp.1731-1736
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• 2009

Recent studies determined that a chronic western-style diet increased the endogenous small heterodimer partner (SHP) protein levels in mice. In experiments with cell cultures, chenodeoxy cholic acid (CDCA) treatment increased endogenous SHP protein levels and reduced the degradation rate of exogenously expressed flag-SHP levels in the human hepatoma cell line, HepG2 cells. In addition, bile acid-induced intestinal fibroblast growth factor-19 (FGF-19) increased the half-life of the exogenously expressed SHP when HepG2 cells were transfected with ad-flag-SHP. However, both the expression level and the degradation rate of the endogenous SHP in response to cholic acid and FGF-19 have not been well understood, either in mice or in cultured HepG2 cells. This study examined the effects of cholic acid treatment on the endogenous SHP protein levels in mice and the effects of FGF-19 on the degradation rate of the endogenous SHP protein in HepG2 cells. Mice fed 0.5% cholic acid in normal chow showed an increase in endogenous SHP protein levels during both 12 hr and 24 hr treatment periods as compared to control mice fed only normal chow. In cultured HepG2 cells, treatment with CDCA did not noticeably change the rate of degradation in the endogenous SHP protein from cells not treated with CDCA. Although consistent with the previous studies on the exogenous ad-flag-SHP protein, treatment with FGF-19 significantly decreased the degradation rate of the endogenous SHP protein when HepG2 cells were treated with cyclohexamide. These results suggest that both bile acids and FGF-19 increase the endogenous SHP protein levels in mouse liver and HepG2 cells.

• BMB Reports
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• v.45 no.3
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• pp.183-188
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• 2012

Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins.

• BMB Reports
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• v.44 no.9
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• pp.572-577
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• 2011

Elevated phospholipase D (PLD) expression prevents cell cycle arrest and apoptosis. However, the roles of PLD isoforms in cell proliferation and apoptosis are incompletely understood. Here, we investigated the physiological significance of the interaction between PLD2 and protein kinase CKII (CKII) in HCT116 human colorectal carcinoma cells. PLD2 interacted with the CKII${\beta}$ subunit in HCT116 cells. The C-terminal domain (residues 578-933) of PLD2 and the N-terminal domain of CKII${\beta}$ were necessary for interaction between the two proteins. PLD2 relocalized CKII${\beta}$ to the plasma membrane area. Overexpression of PLD2 reduced CKII${\beta}$ protein level, whereas knockdown of PLD2 led to an increase in CKII${\beta}$ expression. PLD2-induced CKII${\beta}$ reduction was mediated by ubiquitin-dependent degradation. The C-terminal domain of PLD2 was sufficient for CKII${\beta}$ degradation as the catalytic activity of PLD2 was not required. Taken together, the results indicate that the C-terminal domain of PLD2 can regulate CKII by accelerating CKII${\beta}$ degradation in HCT116 cells.

• Asian-Australasian Journal of Animal Sciences
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• v.3 no.1
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• pp.39-46
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• 1990

This experiment was conducted to study the effects of blood-mixed and heat-treated protein feeds on protein degradation in the rumen, flow of protein to the abomasums and availability of undegraded protein in the intestine of sheep in a $4{\times}4$ Latin square design. Soybean oil meal, rapeseed meal, and whole soybean were mixed with fresh swine blood and dried at $140^{\circ}C$ for 2 h. Proportionate disappearance of apparently digested OM in the postrumen for the blood and heat treated protein group was ranged from 43.2 to 50.5% as compared with 28.0% for the unheated soybean oil meal diet. The treated protein supplements were resulted in greater total N and NAN flow passing at the abomasums than untreated soybean oil meal diet was fed. The quantities of undegraded feed N passing at the abomasums for the treated protein diets was approximately twice as high as that of the untreated soybean oil meal diet and the estimated amount of undegraded N of the protein supplement itself was 79.1 to 84.2% as compared with 15% of soybean oil meal.

• Molecules and Cells
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• v.40 no.12
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• pp.897-905
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• 2017

Cellular protein homeostasis is maintained by two major degradation pathways, namely the ubiquitin-proteasome system (UPS) and autophagy. Until recently, the UPS and autophagy were considered to be largely independent systems targeting proteins for degradation in the proteasome and lysosome, respectively. However, the identification of crucial roles of molecular players such as ubiquitin and p62 in both of these pathways as well as the observation that blocking the UPS affects autophagy flux and vice versa has generated interest in studying crosstalk between these pathways. Here, we critically review the current understanding of how the UPS and autophagy execute coordinated protein degradation at the molecular level, and shed light on our recent findings indicating an important role of an autophagy-associated transmembrane protein EI24 as a bridging molecule between the UPS and autophagy that functions by regulating the degradation of several E3 ligases with Really Interesting New Gene (RING)-domains.

• BMB Reports
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• v.35 no.6
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• pp.590-594
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• 2002

To elucidate the effect of gamma-irradiation on the molecular properties of myoglobin, the secondary and tertiary structures, as well as the molecular weight size of the protein, were examined after irradiation at various irradiation doses. Gamma-irradiation of myoglobin solutions caused the disruption of the ordered structure of the protein molecules, as well as degradation, cross-linking, and aggregation of the polypeptide chains. A SDS-PAGE study indicated that irradiation caused initial fragmentation of the proteins and subsequent aggregation, due to cross-linking of the protein molecules. The effect of irradiation on the protein was more significant at lower protein concentrations. Ascorbic acid protected against the degradation and aggregation of proteins by scavenging oxygen radicals that are produced by irradiation. A circular dichroism study showed that an increase of the irradiation decreased the a-helical content of myoglobin with a concurrent increase of the aperiodic structure content. Fluorescence spectroscopy indicated that irradiation increased the emission intensity that was excited at 280 nm.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.10a
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• pp.135.1-135
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• 2003

To elucidate the effect of gamma-irradiation on the molecular properties of hemoglobin, the secondary, tertiary structure, and the molecular weight size of the protein were examined after irradiation at 0.5, 1, 5, and 10 kGy. Gamma-irradiation of hemoglobin solutions caused the disruption of the ordered structure of the protein molecules, as well as degradation, cross-linking, and aggregation of the polypeptide chains. A SDS-PAGE study indicated that irradiation caused initial fragmentation of the proteins and subsequent aggregation due to cross-linking of the protein molecules. The effect of irradiation on the protein was more significant at lower protein concentrations. Ascorbic acid decreased the degradation and aggregation of proteins by scavenging oxygen radicals that were produced by irradiation. A circular dichroism study showed that irradiation decreased the helical content of hemoglobin with a concurrent increase of the aperiodic structure content. Fluorescence spectroscopy indicated that irradiation decreased the emission intensity that was excited at 280 nm.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.1
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• pp.35-42
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• 1998

Whole faba beans (WFB) were dry roasted at different temperatures (110, 130, $150^{\circ}C$) for 15, 30, 45 minutes to determine the optimal heating conditions of time and temperature to increase nutritional value. Ruminant degradation characteristics of crude protein (CP) of WFB were determined by the nylon bag incubation technique in dairy cows fed 60% hay and 40% concentrate. Measured characteristics of crude protein (CP) were soluble (washable) fraction (S), undegradable fraction (U), lag time (T0), potentially degradable fraction (D) and the rate of degradation (Kd) of insoluble but degradable fraction. Based on measured characteristics, percentage bypass crude protein (%BCP) and bypass crude protein (BCP in g/kg) were calculated. Degradability of CP was reduced by dry roasting (p < 0.01). S was reduced rapidly with increasing time and temperature, from 49.0% in the raw WFB (RWFB) to 26.3% in $150^{\circ}C/45$ min. D varied from 50.7% in RWFB to 73.7% in $150^{\circ}C/45^{\prime}$. U varied from 0% in $130^{\circ}C/45^{\prime}$, $150^{\circ}/30^{\prime}$ and $150^{\circ}/45^{\prime}$ to 0.66% in $110^{\circ}/45^{\prime}$ (0.24% for the RWFB). Lag time (T0) varied from 1.58 h in $130^{\circ}C/30^{\prime}$ to 2.40 h in $150^{\circ}C/45^{\prime}$ (1.87 h for RWFB). Kd varied from 24.2% in the $110^{\circ}C/30^{\prime}$ to 4.3% in $150^{\circ}C/45^{\prime}$ (21.4% for the RWFB). Kd was significantly reduced with time and temperature. All these effects resulted in increasing % BCP from 8.9% in the $110^{\circ}C/45^{\prime}$, 11.3% in the RWFB to 43.1% in the $150^{\circ}C/45$. Therefore BCP increased from 31.3 and 39.9 to 148.4 g/kg respectively. Both %BCP and BCP at $150^{\circ}C/45$ increased nearly 4 times over the raw faba beans. The effects of dry roasting temperature and time on %BCP and BCP seemed to be linear up to the highest values tested. Therefore no optimal dry roasting conditions of time and temperature could be determined at this stage. It was concluded that dry roasting was effective in shifting crude protein degradation from rumen to intestine to reduce unnecessary nitrogen (N) loss in the rumen. To determine the optimal treatment, the digestibility of each treatment should be measured in the next trial using mobile bags technique.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1606-1613
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• 2015

This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.