• Title/Summary/Keyword: protein metabolism

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Characterization of a PyrR-deficient Mutant of Bacillus subtilis by a Proteomic Approach (프로테옴 분석에 의한 Bacillus subtilis PyrR 돌연변이체의 특성)

  • Seul, Keyung-Jo;Cho, Hyun-Soo;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.9-19
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    • 2011
  • The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.

Mechanistic target of rapamycin and an extracellular signaling-regulated kinases 1 and 2 signaling participate in the process of acetate regulating lipid metabolism and hormone-sensitive lipase expression

  • Li, Yujuan;Fu, Chunyan;Liu, Lei;Liu, Yongxu;Li, Fuchang
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1444-1453
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    • 2022
  • Objective: Acetate plays an important role in host lipid metabolism. However, the network of acetate-regulated lipid metabolism remains unclear. Previous studies show that mitogen-activated protein kinases (MAPKs) and mechanistic target of rapamycin (mTOR) play a crucial role in lipid metabolism. We hypothesize that acetate could affect MAPKs and/or mTOR signaling and then regulate lipid metabolism. The present study investigated whether any cross talk occurs among MAPKs, mTOR and acetate in regulating lipid metabolism. Methods: The ceramide C6 (an extracellular signaling-regulated kinases 1 and 2 [ERK1/2] activator) and MHY1485 (a mTOR activator) were used to treat rabbit adipose-derived stem cells (ADSCs) with or without acetate, respectively. Results: It indicated that acetate (9 mM) treatment for 48 h decreased the lipid deposition in rabbit ADSCs. Acetate treatment decreased significantly phosphorylated protein levels of ERK1/2 and mTOR but significantly increased mRNA level of hormone-sensitive lipase (HSL). Acetate treatment did not significantly alter the phosphorylated protein level of p38 MAPK and c-Jun aminoterminal kinase (JNK). Activation of ERK1/2 and mTOR by respective addition in media with ceramide C6 and MHY1485 significantly attenuated decreased lipid deposition and increased HSL expression caused by acetate. Conclusion: Our results suggest that ERK1/2 and mTOR signaling pathways are associated with acetate regulated HSL gene expression and lipid deposition.

An evolving integrative physiology: skeleton and energy metabolism

  • Lee, Na-Kyung
    • BMB Reports
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    • v.43 no.9
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    • pp.579-583
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    • 2010
  • The adipocyte-derived hormone leptin regulates appetite and bone mass. Recent research demonstrates that reciprocally, osteoblasts have a role in controlling energy metabolism. Several genes expressed in osteoblasts are involved in this process, and one of them is the Esp gene. The remaining genes regulate Esp gene expression. OST-PTP, the protein name of Esp, regulates the carboxylation of osteocalcin secreted from osteoblasts, thus affecting insulin sensitivity and insulin secretion. This review provides evidence for a novel interpretation of the connection between bone and energy metabolism and expands our understanding of the novel physiology of bone beyond its classical functions.

Proteomic analysis of rice mutants susceptible to Magnaporthe oryzae

  • Ryu, Hak-Seung;Song, Min-Young;Kim, Chi-Yeol;Han, Muho;Lee, Sang-Kyu;Ryoo, Nayeon;Cho, Jung-Il;Hahn, Tae-Ryong;Jeon, Jong-Seong
    • Plant Biotechnology Reports
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    • v.3 no.2
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    • pp.167-174
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    • 2009
  • To identify genes involved in rice Pi5-mediated disease resistance to Magnaporthe oryzae, we compared the proteomes of the RIL260 rice strain carrying the Pi5 resistance gene with its susceptible mutants M5465 and M7023. Proteins were extracted from the leaf tissues of both RIL260 and the mutant lines at 0, 24, and 48 h after M. oryzae inoculation and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis identified eight proteins that were differently expressed between the resistant and susceptible plants (three down- and five up-regulated proteins in the mutants). The down-regulated proteins included a triosephosphate isomerase (spot no. 2210), a 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (no. 3611), and an unknown protein (no. 4505). In addition, the five up-regulated proteins in the mutants were predicted to be a fructokinase I (no. 313), a glutathione S-transferase (no. 2310), an atpB of chloroplast ATP synthase (no. 3616), an aminopeptidase N (no. 3724), and an unknown protein (no. 308). These results suggest that proteomic analysis of rice susceptible mutants is a useful method for identifying novel proteins involved in resistance to the M. oryzae pathogen.

Effects of the Anabolic Steroid, Nandrolone Pheylpropionate, on Growth and Muscle Protein Metabolism in ACTH-treated Rats (ACTH를 투여한 흰쥐에서 아나보릭스테로이드인 Nandrolone Phenylpropionate가 성장한 근육단백질 대사에 미치는 영향)

  • 주종재
    • Journal of Nutrition and Health
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    • v.29 no.8
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    • pp.874-880
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    • 1996
  • The effects of an anabolic steroid, nandrolone phenylpropionate(NPP), on body weight gain and body protein, and muscle protein metabolism were inestigated in adrenocorticotrophic hormone(ACTH)-treated male and female rats. Daily injections of 100ug/day of ACTH for 7-8 days caused a cessation of growth in females and a net loss of body weight in males which were associated with significant reductions in body protein content. However, food intake was not affected by ACTH in either sex. The weight, protein content and fractional rate of protein synthesis, measured in vivo, of gastrocnemius muscle were all significantly reduced in both sexes. NPP at a dose of 4mg/kg body weight prevented the reduction in body weight gain in ACTH-treate females but not in males. However, boy protein content was increased by NPP in both sexes which was associated with increases in the weight, protein content and fractional rate of protein synthesis of gastrocnemius muscle. ACTH treatment caused a marked increase in plasma concentrations of corticosterone in both sexes. NPP suppressed much of the increases in corticosterone concentrations in both sexes. The results of the present study suggest that NPP exerts at least part of its anabolic effect by reducing plasma concentrations of catabolic glucocorticoid hormones, through suppressing the response of the adrenals to ACTH.

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Effects of Bovine Ash and Calcium Phosphate on Calcium Metabolism in Postmenopausal Osteoporosis Model Rats (골다공증 실혐모델 흰쥐의 칼슘대사에 대한 소뼈회분과 인산칼슘의 섭취 효과)

  • 이연숙
    • Journal of Nutrition and Health
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    • v.28 no.5
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    • pp.434-441
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    • 1995
  • The model rats with postmenopausal osteoporosis were comparatively observed with regard to the effects of bovine ash and calcium phosphate on calcium metabolism. The modelling design involved the five week-old week-old female SD-strain rats ovariectomized and fed a low-Ca diet(20% casein, 0.06% Ca and 0.38% P) for three weeks. The rats were divided into five groups, one of which was fed the low-Ca diet(basal), and the rest of which were divided into five groups, one of which was fed the low-Ca diet(basal), and the rest of which were fed four kinds of Ca-supplemental diets(20% protein, 1.06% Ca and 0.8% P) for three weeks. The Ca-suplements diets contained two kinds of Ca sources, bovine bone ash(BBA) or calcium phosphate, tribasic [Ca3(PO4)2] and two kinds of protein sources, casein or isolated soy protein(ISP). The model rats of postmenopausal osteoporosis fed basal diet showed a significant decrease in Ca utilization in reference to serum Ca concentration, breaking force of bone, Ca and P contents of bone, and Ca absorption and retention. However, the supply of Ca for three weeks demonstrated the improved utilization of Ca. One step further, BBA was more effective than calcium phosphate in improving Ca utilization in ISP-fed groups. On the other hand, no significant difference was seen in casein-fed groups. It is to conclude that BBA could be more effective in accelerating Ca utilization under vulnerable dietary or physiological conditions such as vegetable protein intake and osteoprosis.

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Effect of Fruiting Body of Cordyceps Militaris on Growth, Lipid and Protein Metabolism, and Enzyme Activities in Male Rats (동충하초 자실체가 흰쥐가 성장률, 지질과 단백질 대사 및 효소활성에 미치는 영향)

  • 고진복
    • Journal of Nutrition and Health
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    • v.34 no.7
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    • pp.741-747
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    • 2001
  • The effects of fruiting body of Cordyceps militaris on the growth, the food intake, the food efficiency ratio, the lipid metabolism, the serum protein level ad enzyme activity in male rats were studied. Sprague-Dawley rats were fed four types of diets for five weeks, respectiely: a control diet, a control diet supplemented with 2%, 3% or 4% fruiting body of Cordyceps militaris(CF) powder. In rats fed 2% or 3% CF diets the body weight gain, the food intake, the concentratons of hepatic triglyceride and serum LDL-cholesterol, the atherogenic index, and the total lipid, total cholesterol, triglyceride and phospholipid in serum were similar to those in rats fed the control diet. Whereas, in 4% CF diet these were significantly decrased. But the all CF diets feeding could not decrease the food efficiency ratio, the weights of liver, pancreas, spleen, kidney and heart, and the concentration of serum HDL-cholesterol. Also, it was shown that the concentrations of glucose, total protein, albumin, urea and creatinine, and the GOT, GPT, LDH, ${\gamma}$-GTP and ALP activities were the same levels in serum of rats of fed all the experimental diets.

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Effects of Protein Kinase Inhibitors on In Vitro Protein Phosphorylation and on Secondary Metabolism and Morphogenesis in Streptomyces coelicolor A3(2)

  • Hong, Soon-Kwang;Sueharu, Horinouchi
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.325-332
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    • 1998
  • In vitro phosphorylation experiments with a cell extract of Streptomyces coelicolor A3(2) M130 in the presence of ${\gamma}-[^32P]$]ATP revealed the presence of multiple phosphorylated proteins, including the AfsR/AfsK kinases which control the biosynthesis of A-factor, actinorhodin, and undecylprodigiosin. Phosphorylation of AfsR by a cell extract as an AfsK source was significantly inhibited by Ser/Thr protein kinase inhibitors, staurosporine and K-252a, at concentrations giving 50% inhibition ($IC_50$) of $1{\mu}M\;and\;0.1{\mu}M$, respectively. Further in vitro experiments with the cell extracts showed that phosphorylation of multiple proteins was inhibited by various protein kinase inhibitors with different inhibitory profiles. Manganese and calcium ions in the reaction mixture also modulate phosphorylation of multiple proteins. Manganese at 10 mM greatly enhanced the phosphorylation and partially circumvented the inhibition caused by staurosporine and K-252a. A calcium-activated protein kinase(s) was little affected by these inhibitors. Herbimycin and radicicol, which are known as tyrosine kinase inhibitors, did not show any significant inhibition of AfsR phosphorylation. Consistent with the in vitro effect of the kinase inhibitors, they inhibited aerial mycelium formation and pigmented antibiotic production on solid media. On the contrary, when assayed in liquid culture, the amount of actinorhodin produced was increased by staurosporine and K-252a and greatly decreased by manganese. All of these data clearly show that the genus Streptomyces possesses several protein kinases of eukaryotic types which are involved in the regulatory network for morphogenesis and secondary metabolism.

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Effect of Different Kinds of Protein Fed with Coffee and/or Methionine on the Protein & Lipid Metabolism of Rats (단백질의 종류를 달리한 식이에 첨가한 Coffee 와 Methionine이 흰쥐의 단백질과 지방대사에 미치는 영향)

  • 김영심
    • Journal of Nutrition and Health
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    • v.19 no.4
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    • pp.224-232
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    • 1986
  • This study was performed to investigate the effects of different kinds of dietary protein [plant protein ; Isolated Soy Protein(ISP), animal protein ; casein] on protein and lipid metabolism in rats fed with coffee and/or methionine in diet during four weeks of growing period after weanling. Forty male growing rats fo Sprague-Dawley strain, weighing 92.5$\pm$1.8g, were distributed into 8 groups by randomized complete block design, and fed diets containing 15% of protein by weight either as ISP or casein and 10% ofcalories as corn oil, supplemented with coffee and /or methionine for 4 weeks. Coffee were added at a concentration of 1.4% of diet as instant coffee, and methionine were added to ISP or casein diet to be 0.6% of diet as DL-methionine. Results were followed ; Body weight gain, F.E. R and P.E.R tanded to be higher in methionine added groups than non-methionine groups. The nitrogen content of feces was significantly higher in coffee groups than non-coffee groups, and tended to be higher in ISP groups than casein groups. but was not significantly different with or without methionine. thus, apparent protein digestilbity was significantly lower in coffee groups than non-coffee groups and was significantly lower in ISP groups than cesein groups, but was not significantly different with or without methionine. Total cholesterol content of serum tended to be higher in coffee groups than non-coffee groups, and tended to be lower in methionine groups than non-methionine groups.

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[Retracted] Epinephrine Control of Glycogen Metabolism in Glycogen-associated Protein Phosphatase PP1G/RGLKnockout Mice

  • 김종화;Anna A. DePaoli-Roach
    • BMB Reports
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    • v.35 no.3
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    • pp.283-290
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    • 2002
  • The glycogen-associated protein phosphatase (PP1G/$R_{GL}$) may play a central role in the hormonal control of glycogen metabolism in the skeletal muscle. Here, we investigated the in vivo epinephrine effect of glycogen metabolism in the skeletal muscle of the wild-type and $R_{GL}$ knockout mice. The administration of epinephrine increased blood glucose levels from 200±20 to 325±20 mg/dl in both wild-type and knockout mice. Epinephrine decreased the glycogen synthase -/+ G6P ratio from 0.24±0.04 to 0.10±0.02 in the wild-type, and from 0.17±0.02 to 0.06±0.01 in the knockout mice. Conversely, the glycogen phosphorylase activity ratio increased from 0.21±0.04 to 0.65±0.07 and from 0.30±0.04 to 0.81±0.06 in the epinephrine trated wild-type and knockout mice respectively. The glycogen content of the knockout mice was substantially lower (27%) than that of both wild-type mice; and epinephrine decreased glycogen content in the wild-type and knockout mice. Also, in Western blot analysis there was no compensation of the other glycogen targeting components PTG/R5 and R6 in the knockout mice compared with the wild-type. Therefore, $R_{GL}$ is not required for the epinephrine stimulation of glycogen metabolism, and rather another phosphatase and/or regulatory subunit appears to be involved.