• Title/Summary/Keyword: proteolysis

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Environment-Sensitive Ectodomain Shedding of Epithin/PRSS14 Increases Metastatic Potential of Breast Cancer Cells by Producing CCL2

  • Jang, Jiyoung;Cho, Eun Hye;Cho, Youngkyung;Ganzorig, Binderya;Kim, Ki Yeon;Kim, Moon Gyo;Kim, Chungho
    • Molecules and Cells
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    • v.45 no.8
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    • pp.564-574
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    • 2022
  • Epithin/PRSS14 is a membrane serine protease that plays a key role in tumor progression. The protease exists on the cell surface until its ectodomain shedding, which releases most of the extracellular domain. Previously, we showed that the remaining portion on the membrane undergoes intramembrane proteolysis, which results in the liberation of the intracellular domain and the intracellular domain-mediated gene expression. In this study, we investigated how the intramembrane proteolysis for the nuclear function is initiated. We observed that ectodomain shedding of epithin/PRSS14 in mouse breast cancer 4T1 cells increased depending on environmental conditions and was positively correlated with invasiveness of the cells and their proinvasive cytokine production. We identified selenite as an environmental factor that can induce ectodomain shedding of the protease and increase C-C motif chemokine ligand 2 (CCL2) secretion in an epithin/PRSS14-dependent manner. Additionally, by demonstrating that the expression of the intracellular domain of epithin/PRSS14 is sufficient to induce CCL2 secretion, we established that epithin/PRSS14-dependent shedding and its subsequent intramembrane proteolysis are responsible for the metastatic conversion of 4T1 cells under these conditions. Consequently, we propose that epithin/PRSS14 can act as an environment-sensing receptor that promotes cancer metastasis by liberating the intracellular domain bearing transcriptional activity under conditions promoting ectodomain shedding.

Comparison of Meat Quality Characteristics and Proteolysis Trends associated with Muscle Fiber Type Distribution between Duck Pectoralis Major and Iliotibialis Muscles

  • Cheng, Huilin;Song, Sumin;Park, Tae Sub;Kim, Gap-Don
    • Food Science of Animal Resources
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    • v.42 no.2
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    • pp.266-279
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    • 2022
  • This study was conducted to evaluate the proteolysis trends and change in meat quality during 10 days of cold storage in duck M. pectoralis major (PM) and M. iliotibialis (IL). Duck IL had a higher pH and greater degree of lightness but lower cooking loss than PM (p<0.05). During the 10-day cold storage, the pH value of PM declined significantly (p<0.05), while the meat quality traits of IL were not affected by cold storage (p>0.05). In PM, the redness increased from day 1 to day 5, while cooking loss was lower on day 10 compared to day 5 (p<0.05). There were no significant differences in the activities of cathepsin B and proteasome 20S during cold storage (p>0.05). The activity of calpains declined gradually during 10 days of storage (p<0.05), and the activity of calpains in PM was higher than that in IL (p<0.05). A total of 5,155 peptides were detected and derived from 34 proteins of duck PM muscle, whereas 4,222 peptides derived from 32 proteins were detected from duck IL muscle. Duck PM muscle was composed only of fast type of muscle fiber, whereas IL muscle was composed of both slow and fast types. The proteins responsible for glycolysis or myofibrillar proteins were closely related to changes in meat color or water-holding capacity during cold storage. These results suggest that changes in meat quality characteristics during cold storage are closely related to protein degradation, which is also related to the distribution of muscle fiber types.

Changes in Properties of Deer Antler by Proteolysis and Extraction Conditions (녹용의 단백질가수분해 및 추출조건에 따른 특성 변화)

  • Kim, Jae-Hwa;Yoo, Cheol-Jae;Sin, Kyung-A;Jang, Se-Young;Park, Nan-Young;Jeong, Yong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.89-93
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    • 2011
  • This study was conducted to investigate the proteolysis and extraction conditions of deer antler for application of food materials. ProteAX (A) was the most effective enzyme for proteolysis of deer antler and the proteolysis condition was 0.5% (w/w) for enzyme concentration and 5 hr for proteolysis time. The effect of mixing enzyme ProteAX (A)+KFEN 2 (C) treatment in $60^{\circ}C$, 5 hr was investigated; soluble solid and protein content were the highest with A 0.5% (w/w) and B 0.5% (w/w) concentration. Result for DAH (deer antler hydrolysate) and DA (deer antler) prepared with extraction in $95^{\circ}C$ atmospheric pressure (AP, 6~18 hr) and extraction under $120^{\circ}C$ pressure condition (UP, 15~60 min) after hydrolysis on preceding established condition descriptions indicated that difference in pH according to enzyme treatment and extraction conditions was not significant. Sugar content of DA was $1.5^{\circ}Brix$, DA-UP (under pressure) and DAH-AP (atmospheric pressure) were $2.2^{\circ}Brix$; the highest sugar content of $2.7^{\circ}Brix$ was observed in DAH-UP for 60 min extraction. Also total free sugar, crude protein and collagen content were the highest in DAH-UP for 60 min recording at 1.97%, 742.7 mg/100 g and 498.8 mg/100 g, respectively. From these results, deer antler hydrolysate prepared with extraction under pressure was the most effective for functional characteristics enhancement. Hereafter, various practical uses of materials with enhanced characteristics of antler is expected.

Transmucosal Delivery of Luteinizing Hormone-Releasing Hormone: Effect of Medium Chain Fatty Acid Salts on Stabilization of LHRH in Mucosal Homogenates in vitro. (황체호르몬 유리호르몬의 경점막 수송: 가토 점막균질액 중에서 중쇄지방산염의 LHRH에 대한 안정화 효과)

  • Han, Kun;Park, Jeong-Sook
    • YAKHAK HOEJI
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    • v.38 no.1
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    • pp.67-77
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    • 1994
  • In order to investigate the feasibility of transmucosal delivery of the model peptide, LHRH, metabolism of LHRH and inhibition effect of medium chain fatty acid salts were studied in rabbit mucosal homogenate. LHRH incubated in homogenates of rectal(RE), nasal(NA) and vaginal(VA) mucosa were assayed by HPLC. Five to six degradation products of LHRH were deterted and the degradation of LHRH$(500\;{\mu}g/ml)$ followed the first order kinetics. The main degradation products were found as $LHRH^{1-5}(M-I)$, $LHRH^{1-3}(M-II)$ and $LHRH^{1-6}(M-III)$ by the method of amino acid analysis. The half-lives of LHRH in the mucosal homogenates were found to be less than 20 min at protein concentration of 2.5 mg/ml with the order of VA>NA>RE mucosal homogenate. Medium chain fatty acid salts such as sodium caprylate $(C_8)$, sodium caprate $(C_{10})$ and sodium laurate $(C_{12})$ at the concentration of $0.5%{\sim}1.0%$ inhibit the proteolysis of LHRH significantly. The addition of sodium laurate(0.5%) into the NA and VA mucosal homogenates protected LHRH completely from the degradation.

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Transition of Silk Fibroin by Enzymatic Reaction (효소반응에 의한 견피브로인의 전이)

  • Kim, Dong-Keon;Choi, Jin-Hub;Konishi, Takashi
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.73-78
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    • 1997
  • The crystalline fraction of silk fibroin (Fcp) was obtained from the aqueous solution of silk fibroin hydrolyzed by $\alpha$-chymotrypsin. The molecular weight of Fcp was found approximately 7000 by using high speed GPC. On the other hand, a high molecular weight of PIFcp product could be obtained by the reverse reaction of enzymatic proteolysis of Fcp precipitates. Some parts of this PIFcp have the molecular weights of approximately 17000 and 24000. As a result of x-ray diffraction analysis, the crystal structure of Fcp and PIFcp was turned out silk-II type and silk-I type, respectively. Upon the reverse reaction of enzymatic protelysis, the structural transition occured from silk-II type to silk-I type crystal for the most of Fcp precipitates. It was confirmed that PIFcp might be somewhat stable crystal structure of silk-I type according to the thermal analysis as well as x-ray diffraction method.

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Post-Translational Regulation of miRNA Pathway Components, AGO1 and HYL1, in Plants

  • Cho, Seok Keun;Ryu, Moon Young;Shah, Pratik;Poulsen, Christian Peter;Yang, Seong Wook
    • Molecules and Cells
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    • v.39 no.8
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    • pp.581-586
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    • 2016
  • Post-translational modifications (PTMs) of proteins are essential to increase the functional diversity of the proteome. By adding chemical groups to proteins, or degrading entire proteins by phosphorylation, glycosylation, ubiquitination, neddylation, acetylation, lipidation, and proteolysis, the complexity of the proteome increases, and this then influences most biological processes. Although small RNAs are crucial regulatory elements for gene expression in most eukaryotes, PTMs of small RNA microprocessor and RNA silencing components have not been extensively investigated in plants. To date, several studies have shown that the proteolytic regulation of AGOs is important for host-pathogen interactions. DRB4 is regulated by the ubiquitin-proteasome system, and the degradation of HYL1 is modulated by a de-etiolation repressor, COP1, and an unknown cytoplasmic protease. Here, we discuss current findings on the PTMs of microprocessor and RNA silencing components in plants.

Structural and Functional Relationship of the Catalytical Subunit of Recombinant Pyruvate Dehydrogenase Phosphatase (rPDPc): Limited Proteolysis (Pyruvate dehydrogenase phosphatase의 catalytical subunit의 구조와 활성에 대한 연구)

  • Kim, Young-Mi
    • Environmental Analysis Health and Toxicology
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    • v.17 no.1
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    • pp.73-80
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    • 2002
  • Pyruvate dehydrogenase phosphatase (PDP)와 kinase는 당대사시 해당과정에서의 대사 산물인 pyruvate를 acetyl CoA로 만들어 구연산 회로로 진입시켜 주는 효소인 pyruvate dehydrogenase complex (PDC)의 활성을 조절하는 중요한 효소이다. PDP의 catalytic subunit는 PDC의 dihydrolipoamide acetyltransferase (E2), PDP regulatory subunit (PDPr), 그리고 칼슘 결합 도메인 등으로 구성되어 있는 것으로 추측되어지고 있다. 본 연구에서는 그 구조와 기능과의 상관관계를 알아보기 위해 PDPc를 E. coli JM101에서 발현시켜 순수 정제 후 단백분해 효소를 이용한 제한적 가수분해 방법을 이용해 그 구조와 기능과의 상관관계에 대해 연구하고자 하였다 정제된 PDPc는 trypsin, chymotrypsin, Arg-C 그리고 elastase를 이용하여 3$0^{\circ}C$ 그리고 pH 7.0에서 제한적으로 분해시켰으며 각 분해산물의 아미노 말단의 아미노산 배열을 분석하였다. 그 결과 PDPc는 trypsin, chymotrypsin, elastase에 의해 N-terminal의 50 kD과 C-terminal의 10 kD의 두개의 분해산물을 만들었으며, Arg-C에 의해 50kD의 분해산물은 약 35kD와 15kD으로 더 가수분해가 되었다. 이러한 결과로 볼 때 PDPc는 앞에서 추측한데로 세개의 주요한 기능적 도메인으로 이루어져 있음을 알 수 있었다 또한 C-terminal의 10kD은 PDPc의 활성에는 영향을 주지 않는 것으로 밝혀졌으나 다른 도메인의 기능은 더 연구가 되어져야 할 것으로 생각된다.

Rpn10p is a Receptor for Ubiquitinated Gcn4p in Proteasomal Proteolysis

  • Seong, Ki Moon;Baek, Je-Hyun;Ahn, Byung-Yoon;Yu, Myeong-Hee;Kim, Joon
    • Molecules and Cells
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    • v.24 no.2
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    • pp.194-199
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    • 2007
  • GCN4 is a typical eukaryotic transcriptional activator that is implicated in the expression of many genes involved in amino acids and purine biosyntheses under stress conditions. It is degraded by 26S proteasomes following ubiquitination. However, the immediate receptor for ubiquitinated Gcn4p has not yet been identified. We investigated whether ubiquitinated Gcn4p binds directly to Rpn10p as the ubiquitinated substrate receptor of the 26S proteasome. We found that the level of Gcn4p increased in cells deleted for Rpn10p but not in cells deleted for RAD23 and DSK2, the other ubiquitinated substrate receptors and, unlike Rpn10p, neither of these proteins recognized ubiquitinated Gcn4p. These results suggest that Rpn10p is the receptor that binds the polyubiquitin chain during ubiquitin-dependent proteolysis of Gcn4p.