• Title/Summary/Keyword: cysteine-rich protein 2

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Resveratrol Up-regulates Cysteine-rich Angiogenic Inducer 61 (CYR61) in Human Colorectal Cancer Cells (Resveratrol에 의한 cysteine-rich angiongenic inducer 61 (CYR61) 유전자의 과대발현 조절)

  • Kwak, Eun-Hee;Kim, Jong-Sik
    • Journal of Life Science
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    • v.23 no.2
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    • pp.207-212
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    • 2013
  • In this paper, we investigated whether resveratrol could induce the expression of cysteine-rich angiogenic inducer 61 (CYR61), which is a member of the CCN families. We showed that resveratrol up-regulated CYR61 protein expression in three different human colorectal cancer cell lines. In addition, resveratrol induced CYR61 protein expression in a dose- and time-dependent manner in a HCT116 cell line. To investigate the relationship between various biological activities of resveratrol and CYR61 expression, HCT116 cells were incubated with several NSAIDs, antioxidants, or resveratrol. Interestingly, resveratrol only induced CYR61 protein expression. The expression of CYR61 was not related to the presence of p53. A promoter assay revealed that the 786-bp promoter region (-732/+54) contains a regulatory region and that indole-3-carbinol and 6-gingerol could not induce CYR61 expression. In conclusion, our results indicate up-regulation of CYR61 is extremely resveratrol specific. The results can help to shed light on the unique biological function of resveratrol.

Characterization of Activator of Photopigment and puc Expression, AppA from Rhodobacter sphaeroides 2.4.1

  • Yun, Sang-Hee;Cho, Seung-Hyun;Sa-Ouk kang
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.50-50
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    • 2001
  • Rhodobacter sphaeroides 2.4.1 is a facultatively photoheterotrophic bacterium. The AppA protein is required for increased photo system gene expression upon transition from aerobic respiration to anaerobic photosynthesis condition. This protein has FAD binding domain in amino terminus and cysteine-rich motif in carboxy terminus.(omitted)

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Characterization of the Interaction of Sulfiredoxin (Srx1) with a Vacoular Protein $\alpha$-Mannosidase (Ams1) in Saccharomyces cerevisiae (설피리독신과 알파-만노시다제 간의 단백질 결합 특성에 관한 고찰)

  • Barando, Karen P.;Kim, Il-Han
    • The Journal of Natural Sciences
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    • v.17 no.1
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    • pp.13-29
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    • 2006
  • Most redox-active proteins have thiol-bearing cysteine residues that are sensitive to oxidation. Cysteine thiols oxidized to sulfenic acid are generally unstable, either forming a disulfide with a nearby thiol or being further oxidized to a stable sulfinic acid, which have been viewed as an irreversible protein modification. However, recent studies showed that cysteine residues of certain thiol peroxidases (Prxs) undergo reversible oxidation to sulfinic acid and the reduction reaction is catalyzed by sulfiredoxin (Srx1). Specific Cys residues of various other proteins are also oxidized to sulfinic acid ($Cys-So_2H$). Srxl is considered one of the oxidant proteins with a role in signaling through catalytic reduction of oxidative modification like in the reduction of glutathionylation, a post-translational, oxidative modification that occurs on numerous proteins. In this study, the role of sulfiredoxin in cellular processes, was investigated by studying its interaction with other proteins. Through the yeast two-hybrid system (Y2HS) technique, we have found that Ams1 is a potential and novel interacting protein partner of Srxl. $\alpha$-mannosidase (Ams1) is a resident vacuolar hydrolase which aids in recycling macromolecular components of the cell through hydrolysis of terminal, non-reducing $\alpha$-D-mannose residues. It forms an oligomer in the cytoplasm and under nutrient rich condition and is delivered to the vacuole by the Cytoplasm to Vacuole (Cvt) pathway. Aside from the role of Srxl as a catalyst in the reduction of cysteine sulfenic acid groups, it may play a completely new function in the cellular process as indicated by its interaction with Ams1 of the yeast Saccharomyces cerevisiae.

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Effect of uterine histotroph on embryo development in pigs

  • Han, Hye-In;Lee, Sang-Hee;Song, Eun-Ji;Lee, Seunghyung;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.199-205
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    • 2016
  • The aim of this study was to investigate the effect of uterine histotroph on embryo development and the expression of cysteine-rich protein 2 (CRP2), coatomer subunit gamma-2 (G2COP), myoglobin (MYG), vascular endothelial growth factor D (VEGFD), collagen alpha 4 chain (COL4) and galactoside 3-L-fucosyltransferase 4 (FUT4) proteins in porcine embryo during pre-implantation. Uterine histotroph (UH) was collected from uterine horn on corpus albican phase, and embryos were cultured in porcine zygote medium with UH for 168 hours. Cleavage and blastocyst formation of embryo were detected at 168 hours after in vitro fertilization. And CRP2, G2COP, MYG, VEGFD, COL4 and FUT4 proteins were observed using confocal laser microscope. In results, embryo cleavage rate was not significantly changed by UH, but blastocyst rate was significantly (P<0.05) decreased in UH-treated embryos. Moreover, CRP2, G2COP, MYG, VEGFD, COL4 and FUT4 proteins were expressed in blastomere. CRP2 in embryo was significantly overexpressed (P<0.05), but not G2COP, MYG, VEGFD, COL4 and FUT4 proteins. In summary, UH on corpus albican phase was increased CRP2 protein in embryo, and inhibited blastocyst formation in preimplantation porcine embryos, suggesting that CRP2 may play an interrupter on embryo development in pigs.

Complementation of E. coli cysQ Mutant with Arabidopsis AHL Gene Encoding a 3'(2'),5'-Bisphosphate Nucleotidase

  • Cheong, Jong-Joo;Kwon, Hawk-Bin
    • Journal of Applied Biological Chemistry
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    • v.48 no.4
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    • pp.183-186
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    • 2005
  • Arabidopsis AHL gene encodes a 3'(2')-phosphoadenosine 5'-phosphate (PAP)-specific phosphatase that plays a role in the sulfate activation pathway. We complemented E. coli cysQ mutant defective in cysteine biosynthesis with the AHL gene. AHL cDNA was cloned into the prokaryotic expression vector pKK388-1 and transformed into the bacterial mutant. Since cysQ mutant is a leaky cysteine auxotroph only under aerobic conditions, the bacteria were grown in liquid media with vigorous shaking to provide more aeration. In cysteine-free medium, cysQ mutant and the mutant harboring empty vector did not grow well, whereas cells harboring AHL cDNA exhibited significantly improved growth with doubling time of approximately 3 h. cysQ is known to encode a 3'(2'),5'-diphosphonucleoside 3'(2')-phosphohydrolase (DPNPase). However, our data suggest that cysQ protein has PAP-specific phosphatase activity in addition to DPNPase activity. Microbial complementation procedure described in this paper is useful for structure-activity studies of PAP-specific phosphatases identified from microbes and plants.

A Simple and Rapid Method to Isolate Low Molecular Weight Proteinase Inhibitors from Soybean

  • Krishnan Bari B.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.342-348
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    • 2004
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the $60\%$ isopropanol extract of soybean(Glycine max [L.] Merr.) seed revealed two abundant proteins with molecular masses of 19 and 10 kDa. Amino acid analysis revealed that the isopropanol-extractable protein fraction was rich in cysteine. Two-dimensional gel electro-phoretic analysis indicated that the 19kDa and 10kDa proteins had pI of 4.2 and 4.0 respectively. Peptide mass fingerprints of trypsin digests of the two proteins obtained using matrix-assisted, laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy revealed the 19kDa protein was Kunitz trypsin inhibitor and the 10kDa protein was Bowman-Birk proteinase inhibitor. When resolved under non-denaturing conditions, the isopropanol-extracted proteins inhibited trypsin and chymotrypsin activity. Results presented in this study demonstrate that isopropanol extraction of soybean seed could be used as a simple and rapid method to obtain a protein fraction enriched in Kunitz trypsin and Bowman-Birk proteinase inhibitors. Since proteinase inhibitors are rich in sulfur amino acids and are putative anticarcinogens, this rapid and inexpensive isolation procedure could facilitate efforts in nutrition and cancer research.

Protein and Amino Acid Compositions in Echiurid and Sea Hare Muscles (개불과 군소육의 단백질 및 아미노산 조성)

  • CHOI Yeung-Joon;HAN Young-Sil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.550-556
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    • 1985
  • In order to evaluate the marine mollusc muscle as foodstuff not only from the biochemical aspect but also from the view point of food science, we have analyzed the protein and amino acid compositions of the echiurid (Urechis unicinctus) and sea hare (Aplysia kurodai) muscle. The protein quality of the muscles was also investigated using in vitro methods based on in vitro digestibility, predicted digestibility, computed PER (C-PER) and discriminant computed PER(DC-PER). The remarkable feature of the protein compositions of the both muscles was that water soluble protein occupied a large amount of the muscle protein with fairly lower contents of the salt soluble protein. From the analysis of SDS-PAG electrophoresis, the sarcoplasmic proteins in the echiurid and the sea hare muscles were composed of 15 and 10 subunits, respectively. The free amino acid compositions of the total amino acids in the echiurid and sea hare muscle were characterized with $75\%$ of glycine and alanine, and with $78\%$ of taurine, respectively. The amino acid anaylsis of both muscle proteins showed that the echiurid muscle was rich in glycine, aspartic acid, glutamic acid, arginine and lysine, but was poor in cysteine, while the sea hare muscle was rich in glycine, glutamic acid, aspartic acid and arginine, but was negligible in cysteine and tryptophan. In the total amino acid profiles of the freeze dried muscles in echiurid and sea hare, there was not found a significant difference compared to the amino acid compositions of the muscle proteins. Predicting the protein quality of the echiurid and sea hare muscles using the in vitro method, it was apparently low compared to the muscle protein of fishes.

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Identification and characterization of a rice blast fungal elicitor-inducible Oshin1 gene

  • Kim, Cha-Young;Lee, Sung-Ho
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.45-52
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    • 2009
  • In order to understand the molecular interactions that occur between rice and the rice blast fungus during infection, we previously identified a number of rice blast fungal elicitor-responsive genes from rice (Oryza sativa cv. Milyang 117). Here, we report the cloning and characterization of the rice fungal elicitor-inducible gene Oshin1 (GenBank Accession Number AF039532). Sequence analysis revealed that the Oshin1 cDNA is 1067 bp long and contains an open reading frame encoding 205 amino acid residues. The Oshin1 gene shows considerable sequence similarity to the tobacco hin1 and hin2 genes. The predicted Oshin1 protein has a cysteine-rich domain at the N-terminus and is rich in leucine, serine, and alanine residues. Southern blot analysis suggests that Oshin1 gene is a member of a small gene family in the rice genome. To examine the expression of Oshin1, Northern blot analysis was conducted. Expression of the Oshin1 transcript is rapidly induced in suspension-cultured rice cells treated with fungal elicitor, salicylic acid or hydrogen peroxide. In addition, Oshin1 transcript levels are rapidly increased by treatment with $Ca^{2+}$/A23187. The expression of Oshin1 was also elevated in 3-week old leaf tissues upon ethephon application or fungal elicitor treatment. Our results suggest that the Oshin1 gene is involved in plant defense responses to environmental stresses.

Analysis of brain protein expression in developing mouse fetus (임신일령에 따른 생쥐 태아 뇌조직의 단백질 발현 양상 분석)

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yun-Fei;Woo, Je-Seok;Jin, Dong-Il
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.65-70
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    • 2011
  • Development of mouse fetus brains can be defined morphologically and functionally by three developmental stages, embryo day (ED) 16, postnatal stage one week and eight weeks. These defined stages of brain development may be closely associated with differential gene expression rates due to limited cellular resources such as energy, space, and free water. Complex patterns of expressed genes and proteins during brain development suggests the changes in relative concentrations of proteins rather than the increase in numbers of new gene products. This study was designed to evaluate early protein expression pattern in mouse fetus brain. The mouse brain proteome of fetus at ED 15.5, and 19.5 was obtained using 2-dimensional gel electrophoresis (DE). Analysis of the 2-DE gels in pH 3-10 range revealed the presence of 15 differentially expressed spots, of which 11 spots were identified to be known proteins following MALDI-TOF analysis; 3 spots were up-regulated and 8 spots were down-regulated in the mouse fetus brain at ED 15.5. UP-regulated proteins were identified as MCG18238, isoform M2 of pyruvate kinase isozymes M1/M2, isoform 2 of heterogeneous nuclear ribonucleoprotein K, heterogeneous nuclear ribonucleoprotein H2, creatine kinase B-type, 40S ribosomal protein SA and hemoglobin subunit beta-H1. Down-regulated proteins were putative uncharacterized protein, lactoylglutathione lyase and secreted acidic cysteine rich glycoprotein. Our results revealed composite profiles of mouse fetus brain proteins related to mouse fetus development by 2-DE analysis implying possible roles of these proteins in neural differentiation.

Baicalein Inhibits Epithelial to Mesenchymal Transition via Downregulation of Cyr61 and LOXL-2 in MDA-MB231 Breast Cancer Cells

  • Nguyen, Linh Thi Thao;Song, Yeon Woo;Cho, Somi Kim
    • Molecules and Cells
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    • v.39 no.12
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    • pp.909-914
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    • 2016
  • Epithelial-mesenchymal transition (EMT) is a critical step in the acquisition of the migratory and invasive capabilities associated with metastatic competence. Cysteine-rich protein 61 (CCN1/Cyr61) has been implicated as an important mediator in the proliferation and metastasis of breast cancer. Hence, Cyr61 and associated pathways are attractive targets for therapeutic interventions directed against the EMT. In the present study, we report that baicalein significantly inhibits the expression of Cyr61 and migration and invasion of MDA-MB231 human breast cancer cells. Exposure to baicalein led to increased E-cadherin expression, possibly due to the ubiquitination of Snail and Slug, which was mediated by the Cyr61/Akt/glycogen synthase kinase $3{\beta}$ ($GSK3{\beta}$) pathway. Further analysis revealed that baicalein inhibited the expression of lysyl oxidase like-2 (LOXL-2), which is a functional collaborator of Snail and Slug, and subsequently attenuated the direct interaction between LOXL-2 and Snail or Slug, thereby enhancing $GSK3{\beta}$-dependent Snail and Slug degradation. Our findings provide new insights into the antimetastatic mechanism of baicalein and may contribute to its beneficial use in breast cancer therapies.