• 한국축산식품학회지
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• 제38권5호
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• pp.1029-1042
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• 2018

This study aimed to investigate the use of golden thread extract (GTE), clove extract (CE), and commercially available nitrite for retarding lipid and protein oxidation and for maintaining color stability and sensory attributes in beef patties stored at $4^{\circ}C$. GTE, CE, and nitrite treatment samples were found to be efficient in retarding lipid oxidation as all three treatments resulted in low thiobarbituric acid reactive substance (TBARS) content (p<0.05). By using GTE, CE, and nitrite into beef patties, protein oxidation was not developed. Incorporation of GTE and CE into beef patties maintained color stability by protecting against the decrease of $L^*$, $a^*$, $b^*$, chroma, and hue angle values and exhibited significant influence on sensory characteristics, including color and odor of beef patties (p<0.05). Compared to commercially available nitrite, GTE and CE were more effective as antioxidants for inhibiting lipid oxidation, and preserving color stability of fresh beef patties. The study indicated that GTE and CE could be utilized efficiently to extend the shelf life of beef patties.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.355-359
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• 2006

The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

• Molecules and Cells
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• 제43권4호
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• pp.373-383
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• 2020

Our previous study revealed a novel role of Fas-associated death domain-containing protein (FADD) in islet development and insulin secretion. Insulin-degrading enzyme (IDE) is a zinc metalloprotease that selectively degrades biologically important substrates associated with type 2 diabetes (T2DM). The current study was designed to investigate the effect of FADD phosphorylation on IDE. We found that the mRNA and protein levels of IDE were significantly downregulated in FADD-D mouse livers compared with control mice. Quantitative real-time polymerase chain reaction analysis showed that FADD regulates the expression of IDE at the transcriptional level without affecting the stability of the mRNA in HepG2 cells. Following treatment with cycloheximide, the IDE protein degradation rate was found to be increased in both FADD-D primary hepatocytes and FADD-knockdown HepG2 cells. Additionally, IDE expression levels were reduced in insulin-stimulated primary hepatocytes from FADD-D mice compared to those from control mice. Moreover, FADD phosphorylation promotes nuclear translocation of FoxO1, thus inhibiting the transcriptional activity of the IDE promoter. Together, these findings imply a novel role of FADD in the reduction of protein stability and expression levels of IDE.

• 폴리머
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• 제28권6호
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• pp.502-508
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• 2004

암을 치료하기 위한 양이온성 리포솜은 화학치료요법의 분야에서 발전되어 왔다. 양이온성 리포솜은 음이온성의 세포 표면에 정전기적 상호작용에 의해 결합이 된다. 본 연구의 목적은 음이온성 세포와 이온결합을 할 수 있는 양이온성 리포솜을 제조하는 것이다. 양이온성 리포솜은 지질인 1,2-디스테로일-sn-글리콜-3-포스포에탄올아민 (DSPE)과 양이온성 고분자인 폴리에틸렌이민 (PEI)을 그라프트 중합으로 합성된 물질로부터 제조하였다. 리포솜 표면의 이온 특성은 제타 포텐셜을 측정하여 확인하였다. 소 혈청 수용액에서 리포솜의 혈장 단백질 흡착 특성은 입자 크기와 탁도 변화를 측정하여 확인하였다. 완충 용액 속에서 리포솜 안정성을 예측하기 위하여 입자 크기를 7일 동안 상온에서 측정하였다. 양이온성 리포솜은 소 혈청 수용액에서 많은 양의 혈장 단백질이 흡착되었다. 이는 혈장 단백은 음전하를 가지고 있어서 양이온성 리포솜의 표면과 잘 붙기 때문이다. 양이온성 폴리에틸렌이민을 이용한 리포솜의 표면 변형은 소 혈청 수용액 내에서 단백질 흡착을 강화시킨다는 것을 예상하게 한다. 또한, 제조된 리포솜의 완충 용액 내에서 7일 동안 안정한 상태임을 관찰하였다.

• 한국축산식품학회지
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• 제39권5호
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• pp.768-779
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• 2019

This study was performed to assess the comparison of the effects amongst butylated hydroxytoluene (BHT), clove extract (CE), and ascorbic acid (AA) as antioxidants on the oxidative stability and color values in fresh beef patties. The adding of BHT, AA, and CE to patties significantly restrained lipid oxidation, lowered hue angle as color value, and expanded redness and chroma values of fresh beef patties in comparison to the control (p<0.05). BHT and AA significantly led to impede the protein oxidation of patties by lowering carbonyl content (p<0.05). CE had no negative effect on protein oxidation. The antioxidant effects of BHT, AA, and CE were obviously manifested. Nonetheless, BHT, AA, and CE appeared to have insignificant difference of each other for lowering the protein oxidation at the end of storage. BHT and CE represented lowered lipid oxidation in comparison to AA. The antioxidant effects of BHT, AA, and CE on lipid oxidation were more marked than the effects on protein oxidation. Furthermore, CE as a natural antioxidant evinced the efficiency in oxidative stability and color stability in fresh beef patties. The study implied that CE could substitute the use of BHT and AA when making beef patties during storage.

• Molecules and Cells
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• 제44권2호
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• pp.101-115
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• 2021

The INO80 chromatin remodeling complex has roles in many essential cellular processes, including DNA replication. However, the mechanisms that regulate INO80 in these processes remain largely unknown. We previously reported that the stability of Ino80, the catalytic ATPase subunit of INO80, is regulated by the ubiquitin proteasome system and that BRCA1-associated protein-1 (BAP1), a nuclear deubiquitinase with tumor suppressor activity, stabilizes Ino80 via deubiquitination and promotes replication fork progression. However, the E3 ubiquitin ligase that targets Ino80 for proteasomal degradation was unknown. Here, we identified the C-terminus of Hsp70-interacting protein (CHIP), the E3 ubiquitin ligase that functions in cooperation with Hsp70, as an Ino80-interacting protein. CHIP polyubiquitinates Ino80 in a manner dependent on Hsp70. Contrary to our expectation that CHIP degrades Ino80, CHIP instead stabilizes Ino80 by extending its half-life. The data suggest that CHIP stabilizes Ino80 by inhibiting degradative ubiquitination. We also show that CHIP works together with BAP1 to enhance the stabilization of Ino80, leading to its chromatin binding. Interestingly, both depletion and overexpression of CHIP compromise replication fork progression with little effect on fork stalling, as similarly observed for BAP1 and Ino80, indicating that an optimal cellular level of Ino80 is important for replication fork speed but not for replication stress suppression. This work therefore idenitifes CHIP as an E3 ubiquitin ligase that stabilizes Ino80 via nondegradative ubiquitination and suggests that CHIP and BAP1 act in concert to regulate Ino80 ubiquitination to fine-tune its stability for efficient DNA replication.

• Genomics & Informatics
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• 제19권3호
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• pp.29.1-29.10
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• 2021

In our previous studies, we have demonstrated the association of certain variants of the thyroid-stimulating hormone receptor (TSHR), thyroid peroxidase (TPO), and thyroglobulin (TG) genes with congenital hypothyroidism. Herein, we explored the mechanistic basis for this association using different in silico tools. The mRNA 3'-untranslated region (3'-UTR) plays key roles in gene expression at the post-transcriptional level. In TSHR variants (rs2268477, rs7144481, and rs17630128), the binding affinity of microRNAs (miRs) (hsa-miR-154-5p, hsa-miR-376a-2-5p, hsa-miR-3935, hsa-miR-4280, and hsa-miR-6858-3p) to the 3'-UTR is disrupted, affecting post-transcriptional gene regulation. TPO and TG are the two key proteins necessary for the biosynthesis of thyroid hormones in the presence of iodide and H₂O₂. Reduced stability of these proteins leads to aberrant biosynthesis of thyroid hormones. Compared to the wild-type TPO protein, the p.S398T variant was found to exhibit less stability and significant rearrangements of intra-atomic bonds affecting the stoichiometry and substrate binding (binding energies, ΔG of wild-type vs. mutant: -15 vs. -13.8 kcal/mol; and dissociation constant, K_d of wild-type vs. mutant: 7.2E^-12 vs. 7.0E^-11 M). The missense mutations p.G653D and p.R1999W on the TG protein showed altered ΔG(0.24 kcal/mol and 0.79 kcal/mol, respectively). In conclusion, an in silico analysis of TSHR genetic variants in the 3'-UTR showed that they alter the binding affinities of different miRs. The TPO protein structure and mutant protein complex (p.S398T) are less stable, with potentially deleterious effects. A structural and energy analysis showed that TG mutations (p.G653D and p.R1999W) reduce the stability of the TG protein and affect its structure-functional relationship.

• 한국식품영양과학회지
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• 제28권1호
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• pp.153-160
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• 1999

Storage stability of boiled and freeze dried loach and antioxidative effect of Zanthoxylum schinifolium were studied to confirm the possibility in development of instant choo o tang(Korean traditional loach soup). Packaging and storage temperature did not cause a measurable change in in vitro protein digestibility and trypsin indigestible substrate within 45 days of storage but remarkable quality changes were occurred in all samples stored after 60 days. Vacuum packaging and low temperature storage(4 oC) had some effect in retarding protein quality deterioration due to delaying polyunsaturated fatty acid oxidation. Maximum peroxide value and TBA value were reached in 15 days, and there were a slow(TBA value) and rapid reduction(POV) after peaks were reached. In contrast, increasing brown pigment development and fluorescence intensity continued until 90 days of storage. Treatment of ethanolic extracts from Zanthoxylum schinifolium prior to freeze drying could protect against lipid oxidation of freeze dried loach products.

• 생명과학회지
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• 제21권8호
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• pp.1067-1075
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• 2011

CCDC98 단백질은 BRCA1-A 복합체를 DNA 손상 부위로 이동시킴으로써 DNA손상에 의하여 유도되는 G2/M cell cycle checkpoint에 중요한 역할을 한다고 알려져 있다. 하지만 많은 연구에도 불구하고 CCDC98 단백질의 기능에 대해서 알려진 바가 거의 없다. 본 연구는 CCDC98 단백질의 기능을 밝히고자 tandem affinity purification 방법을 수행하였다. 그 결과 Wilms tumor 1-associating protein (WTAP)을 CCDC98의 결합 단백질로 분리 동정하였다. 이들 단백질의 결합을 in vivo and in vitro binding assays를 통하여 확인하였다. 또한, CCDC98 단백질이 cyclin B1의 발현을 억제함을 확인하였는데, 이는 WTAP 단백질의 발현을 조절함으로써 이루어진다는 것을 확인하였다. 이는 CCDC98 단백질이 새로운 세포주기 조절자라는 것을 증명하는 결과이다.

• Applied Biological Chemistry
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• 제49권4호
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• pp.304-308
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• 2006

누에번데기에 함유되어 있는 불용성 단백질을 가용성 단백질로 추출시키기 위하여 누에번데기에 Bacillus sp. JH-209 균주로부터 생산된 protease를 작용시켰다. 이때 누에번데기 단백질의 추출을 위한 적정 pH는 pH $7{\sim}11$까지의 알칼리 영역에서 추출율의 증가를 보였다. 최적 온도는 $40^{\circ}C$였고, 최적 작용시간은 11시간이었고, 효소의 최적 첨가량은 60 unit 정도였다. 효소처리 된 누에번데기 단백질은 효소처리하지 않은 대조구에 비해 기포력, 기포안정성, 유화력과 유화안정성이 증가하였고, 유지흡착력과 수분흡착력도 대조구에 비해서 높은값을 나타내었다.