• Title/Summary/Keyword: Enzyme regulation

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Activity and application of 20α-hydroxysteroid dehydrogenase in rat 1. Changes in activities of 20α-hydroxysteroid dehydrogenase and serum progesterone concentration (Progesterone의 이화(異化)효소, 20α-hydroxysteroid dehydrogenase의 활성 및 활용에 관한 연구 1. 20α-hydroxysteroid dehydrogenase의 활성 및 혈청 progesterone 농도의 변화)

  • Kang, Chung-boo;Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.34 no.4
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    • pp.867-872
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    • 1994
  • Progesterone catabolizing enzyme, the enzyme $20{\alpha}$-hydroxysteroid dehydrgenase($20{\alpha}$-HSD) is pivotal in the regulation of ovarian function in rodents, which catabolizes progesterone into biologically inactive $20{\alpha}$-hydroxypregn-4-en-3-one($20{\alpha}$-OHP). In this study was carried out the influence of $20{\alpha}$-HSD activity of ovarian function, we investigated changes in ovarian cytosol $20{\alpha}$-HSD activity and serume progesterone concentration during the estrous cycles and pregnancy in rat. During the estrous cycles, the $20{\alpha}$-HSD activities were highest on the progestrous, but serum progesterone concentration was lowest on this phase. During the pregnancy, the $20{\alpha}$-HSD activities were relatively higher early pregnancy(day-1-3 gestation) and late pregnancy(day 21 to parturition), serum progesterone concentration was maintained significantly high to day 19 of gestation. The $20{\alpha}$-HSD activities were lower during the middle pregnancy. From these results, ovarian $20{\alpha}$-HSD activities may possibly act as physiologically very important in the control and maintenance of estrous cycles in rat.

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Overexpression of starch branching enzyme 1 gene improves eating quality in japonica rice

  • Sun, Ming-Mao;Lee, Hye-Jung;Abdula, Sailila E.;Jee, Moo-Geun;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.88-101
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    • 2013
  • Eating quality of rice attracts more and more attention from rice-eating consumers in the recent years. Thus, improvement of eating quality of cooked rice has become one of the most important breeding goals in japonica rice. Here, the generation of transgenic japonica rice with improved eating quality and grain yield are reported. Overexpression of OsSbe1 gene encoding rice starch branching enzyme 1 was driven by 35S promoter. Eleven independent homozygous $T_3$ transgenic lines were characterized and had shown higher palatability (71.2 ~ 72.6) than wild type Gopum (70.4). Moreover, transgenic rice lines showed an increase in 1000-grain weight and number of spikelets per panicle compared with the wild type. The yield of milled rice was 562.8 ~ 596.7 kg/10a in eight $T_3$ lines, but 542.1 kg/10a in wild type. Gene expression analyses in mRNA transcription and enzyme activity levels suggest that improved eating quality is due to the up-regulation of OsSbe1 gene.

Regulation of NAD+- Specific Isocitrate Dehydrogenase from Pythium ultimum

  • Kim, Hak-Ryul;Weete, John D.
    • BMB Reports
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    • v.32 no.4
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    • pp.385-392
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    • 1999
  • The $NAD^+$-specific activity of a dual coenzyme-specific isocitrate dehydrogenase (IDH; EC 1.1.1.41) from the primitive fungus Pythium ultimum was investigated to elucidate the regulatory factors that may influence the intracellular distribution of carbon and the availability of intermediates, e.g. citrate, for fatty acid synthesis. Inhibition of $NAD^+$-IDH activity by diphospho- and triphosphonucleotides (ATP, ADP, and GTP) reflected the sensitivity of this enzyme to cellular energy charge even though monophosphonucleotides (AMP and GMP) had little effect on activity. NADPH, but not NADH, substantially inhibited $NAD^+$-IDH activity, showing noncompetitive inhibition with isocitrate. Oxalacetate and ${\alpha}$-ketoglutarate showed competitive inhibition with isocitrate, while citrate and cis-aconitate showed mixed-noncompetitive inhibition with isocitrate. Inhibition by these substances ranged from 29 to 46% at 10 mM. The inhibitory effect of oxalacetate was increased synergistically by glyoxylate, which alone caused 31% uncompetitive inhibition at 10 mM, and a mixture of the two substances at 1 mM each showed 98% inhibition of $NAD^+$-IDH activity. The regulation of $NAD^+$-IDH in Pythium ultimum seems to be a complex process involving mitochondrial metabolites. The addition of glyoxylate (3 mM) and oxalacetate (3 mM) to the culture medium resulted in the production of 49% more lipid by P. ultimum.

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Antiproliferative Effects of Crocin in HepG2 Cells by Telomerase Inhibition and hTERT Down-Regulation

  • Noureini, Sakineh Kazemi;Wink, Michael
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2305-2309
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    • 2012
  • Crocin, the main pigment of Crocus sativus L., has been shown to have antiproliferative effects on cancer cells, but the involved mechanisms are only poor understood. This study focused on probable effect of crocin on the immortality of hepatic cancer cells. Cytotoxicity of crocin ($IC_{50}$ 3 mg/ml) in hepatocarcinoma HepG2 cells was determined after 48 h by neutral red uptake assay and MTT test. Immortality was investigated through quantification of relative telomerase activity with a quantitative real-time PCR-based telomerase repeat amplification protocol (qTRAP). Telomerase activity in 0.5 ${\mu}g$ protein extract of HepG2 cells treated with 3 mg/ml crocin was reduced to about 51% as compared to untreated control cells. Two mechanisms of inhibition, i.e. interaction of crocin with telomeric quadruplex sequences and down regulation of hTERT expression, were examined using FRET analysis to measure melting temperature of a synthetic telomeric oligonucleotide in the presence of crocin and quantitative real-time RT-PCR, respectively. No significant changes were observed in the $T_m$ telomeric oligonucleotides, while the relative expression level of the catalytic subunit of telomerase (hTERT) gene showed a 60% decrease as compared to untreated control cells. In conclusion, telomerase activity of HepG2 cells decreases after treatment with crocin, which is probably caused by down-regulation of the expression of the catalytic subunit of the enzyme.

Changes of PBP Quantity and FNR Activity by Light Wavelengths in Anabaena variabilis (光波長에 따른 Anabaena variabilis 의 Phycobiliprotein 含量 및 FNR 活性度 變化)

  • Kim, Jung-Suk;Chang, Nam-Kee
    • The Korean Journal of Ecology
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    • v.14 no.1
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    • pp.87-99
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    • 1991
  • Changes of phycobiliproteins(PBP) quantity and ferredoxin-NADP reductase(FNR) activity were investigated in various light illuminated cyanobacteria, Anabaena variabilis. PBP components were increased under blue light illumination, whereas decreased under red light illumination. PBP contents were twofolds in blue light than in red light. In view of the PBP composition, allophycocyanin(APC) in red light was higher 5.5% and phycoerythrocyanin(PEC) in blue light was higher 2.2% than in white light-illuminated PBP. It was suggested that PBP changes in bule light be the results of regulation of photosysthetic efficiency and protection of photosystem, whereas PBP changes in red light be effected by adaptation of adequate harvesting of light energy in photosystem. Changes of FNR activity were highest in red light, and sequenced lower to blue light and green light. It means that light-dependent production rate of NADP is the highest in red light. The difference of values was larger than that of values in comparison of red and blue light. It was suggested that increasing of FNR activity be due not to the function of isozyme, but to the synthesis of enzymes. Because of NAD/NADP regulation-effect to metabolism, it was considered that FNR activity might influence the metabolism indirectly and explain the probability of regulation in pathways of key enzyme activation. FNR activity was directly proportional to intensity of light. Optimum temperature and pH were about 25℃ and 7.5, respectively.

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Src Redox Regulation: There Is More Than Meets the Eye

  • Chiarugi, Paola
    • Molecules and Cells
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    • v.26 no.4
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    • pp.329-337
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    • 2008
  • Src-family kinases are critically involved in the control of cytoskeleton organization and in the generation of integrin-dependent signaling responses, inducing tyrosine phosphorylation of many signaling and cytoskeletal proteins. Activity of the Src family of tyrosine kinases is tightly controlled by inhibitory phosphorylation of a carboxy-terminal tyrosine residue, inducing an inactive conformation through binding with its SH2 domain. Dephosphorylation of C-ter tyrosine, as well as its deletion of substitution with phenylalanine in oncogenic Src kinases, leads to autophosphorylation at a tyrosine in the activation loop, thereby leading to enhanced Src activity. Beside this phophorylation/dephosphorylation circuitry, cysteine oxidation has been recently reported as a further mechanism of enzyme activation. Mounting evidence describes Src activation via its redox regulation as a key outcome in several circumstances, including growth factor and cytokines signaling, integrin-mediated cell adhesion and motility, membrane receptor cross-talk as well in cell transformation and tumor progression. Among the plethora of data involving Src kinase in physiological and pathophysiological processes, this review will give emphasis to the redox component of the regulation of this master kinase.

Tumor necrosis factor α-converting enzyme inhibitor attenuates lipopolysaccharide-induced reactive oxygen species and mitogen-activated protein kinase expression in human renal proximal tubule epithelial cells

  • Bae, Eun Hui;Kim, In Jin;Choi, Hong Sang;Kim, Ha Yeon;Kim, Chang Seong;Ma, Seong Kwon;Kim, In S.;Kim, Soo Wan
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.2
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    • pp.135-143
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    • 2018
  • Tumor necrosis $factor-{\alpha}$ ($TNF{\alpha}$) and the angiotensin system are involved in inflammatory diseases and may contribute to acute kidney injury. We investigated the mechanisms by which $TNF{\alpha}$-converting enzyme (TACE) contributes to lipopolysaccharide (LPS)-induced renal inflammation and the effect of TACE inhibitor treatment on LPS-induced cellular injury in human renal proximal tubule epithelial (HK-2) cells. Mice were treated with LPS (10 mg/kg, i.p.) and HK-2 cells were cultured with or without LPS ($10{\mu}g/ml$) in the presence or absence of a type 1 TACE inhibitor ($1{\mu}M$) or type 2 TACE inhibitor ($10{\mu}M$). LPS treatment induced increased serum creatinine, $TNF{\alpha}$, and urinary neutrophil gelatinase-associated lipocalin. Angiotensin II type 1 receptor, mitogen activated protein kinase (MAPK), and TACE increased, while angiotensin-converting enzyme-2 (ACE2) expression decreased in LPS-induced acute kidney injury and LPS-treated HK-2 cells. LPS induced reactive oxygen species and the down-regulation of ACE2, and these responses were prevented by TACE inhibitors in HK-2 cells. TACE inhibitors increased cell viability in LPS-treated HK-2 cells and attenuated oxidative stress and inflammatory cytokines. Our findings indicate that LPS activates renin angiotensin system components via the activation of TACE. Furthermore, inhibitors of TACE are potential therapeutic agents for kidney injury.

Characterization of Phosphatidylcholine-Hydrolyzing Phospholipase D in the Scuticociliate Parasite, Uronema marinum

  • Seo, Jung-Soo;Kim, Moo-Sang;Kim, Na-Young;Ahn, Sang-Jung;Jee, Bo-Young;Jung, Sung-Hee;Kim, Jin-Woo;Kim, Ki-Hong;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.21 no.1
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    • pp.1-11
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    • 2008
  • We report the existence of new type of phosphatidylcholine-hydrolyzing phospholipase D (PLD), which has been characterized and partially purified in the scuticociliate, Uronema marinum. The enzyme from partial purification showed that it was existed in membrane fraction and was a neutral PLD, which catalyzed both transphosphatidylation and hydrolysis reaction. The activity of partially purified membrane-bound PLD was also found to be optimal at pH 7.0-7.5 for 2 hours at 37℃ and depended strictly on the presence of Ca2+ (2.5 mM) and Mg2+ (1.6 mM). Immunoblot analysis indicated that the enzyme was distinct from hPLD1 (human PLD1) and hPLD2 (human PLD2) because it was not recognized by a polyclonal antibody raised to the 12 terminal amino acid of these enzymes. We also found that the membrane-bound PLD is a PIP2-dependent PLD and that GTP-binding proteins are not implicated in the regulation of this enzyme: This enzyme activity is markedly stimulated by phosphatidylinositol 4,5-bisphosphate (PIP2) but not by the small G-protein Arf and GTPrS. In addition, this enzyme was capable of hydrolyzing phosphatidylcholine (PC) but not phosphatidylethanolamine (PE), implying that PC was a preferred substrate.

Regulation of Ethylene Biosynthesis by Phytohormones in Etiolated Rice (Oryza sativa L.) Seedlings

  • Lee, Han Yong;Yoon, Gyeong Mee
    • Molecules and Cells
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    • v.41 no.4
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    • pp.311-319
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    • 2018
  • The gaseous hormone ethylene influences many aspects of plant growth, development, and responses to a variety of stresses. The biosynthesis of ethylene is tightly regulated by various internal and external stimuli, and the primary target of the regulation is the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), which catalyzes the rate-limiting step of ethylene biosynthesis. We have previously demonstrated that the regulation of ethylene biosynthesis is a common feature of most of the phytohormones in etiolated Arabidopsis seedlings via the modulation of the protein stability of ACS. Here, we show that various phytohormones also regulate ethylene biosynthesis from etiolated rice seedlings in a similar manner to those in Arabidopsis. Cytokinin, brassinosteroids, and gibberellic acid increase ethylene biosynthesis without changing the transcript levels of neither OsACS nor ACC oxidases (OsACO), a family of enzymes catalyzing the final step of the ethylene biosynthetic pathway. Likewise, salicylic acid and abscisic acid do not alter the gene expression of OsACS, but both hormones downregulate the transcript levels of a subset of ACO genes, resulting in a decrease in ethylene biosynthesis. In addition, we show that the treatment of the phytohormones results in distinct etiolated seedling phenotypes, some of which resemble ethylene-responsive phenotypes, while others display ethylene-independent morphologies, indicating a complicated hormone crosstalk in rice. Together, our study brings a new insight into crosstalk between ethylene biosynthesis and other phytohormones, and provides evidence that rice ethylene biosynthesis could be regulated by the post-transcriptional regulation of ACS proteins.

Developmental Roles of D-bifunctional Protein-A Zebrafish Model of Peroxisome Dysfunction

  • Kim, Yong-Il;Bhandari, Sushil;Lee, Joon No;Yoo, Kyeong-Won;Kim, Se-Jin;Oh, Gi-Su;Kim, Hyung-Jin;Cho, Meyoung;Kwak, Jong-Young;So, Hong-Seob;Park, Raekil;Choe, Seong-Kyu
    • Molecules and Cells
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    • v.37 no.1
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    • pp.74-80
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    • 2014
  • The peroxisome is an intracellular organelle that responds dynamically to environmental changes. Various model organisms have been used to study the roles of peroxisomal proteins in maintaining cellular homeostasis. By taking advantage of the zebrafish model whose early stage of embryogenesis is dependent on yolk components, we examined the developmental roles of the D-bifunctional protein (Dbp), an essential enzyme in the peroxisomal ${\beta}$-oxidation. The knockdown of dbp in zebrafish phenocopied clinical manifestations of its deficiency in human, including defective craniofacial morphogenesis, growth retardation, and abnormal neuronal development. Overexpression of murine Dbp rescued the morphological phenotypes induced by dbp knockdown, indicative of conserved roles of Dbp during zebrafish and mammalian development. Knockdown of dbp impaired normal development of blood, blood vessels, and most strikingly, endoderm-derived organs including the liver and pancreas - a phenotype not reported elsewhere in connection with peroxisome dysfunction. Taken together, our results demonstrate for the first time that zebrafish might be a useful model animal to study the role of peroxisomes during vertebrate development.