• Title/Summary/Keyword: Protein phosphatase

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The Effect of Dietary Protein Source and Sulfur Amino acid Content on bone Metabolism in Growing Rats (식이 단백질의 종류와 함황아미노산 함량이 성장기 쥐의 골밀도에 미치는 영향)

  • 최미자;정소형
    • Journal of Nutrition and Health
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    • v.37 no.2
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    • pp.100-107
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    • 2004
  • This study was performed to evaluate the effect of dietary protein source and sulfur amino acid content on bone metabolism in ra. Thirty male rats (body weight 145$\pm$2g) were divided into three groups. The rats in the first group were fed on casein 20% diet as animal protein source and those in the second group were fed on soy 20% diet as plant protein source. Sulfur amino acid ratio of these group was 1.07:1. The rats in the third group were fed on soy 20% diet and the sulfur amino acid were supplemented with the amount contained as much in the soy 20% diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks, The total body, spine, femur bone mineral density and bone mineral content were measured using Dual Energy X-ray Absorptiometry Calcium, phosphate, pyridinoline, creatinine in urine and calcium, phosphate, alkaline phosphatase, osteocalcin in serum were measured. During the experimental period, plant protein (soy protein) group had a lower urinary Ca excretion, urine pyridinoline & crosslinks value and had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein) group. There were no significant differences in serum calcium, phosphate, alkaline phosphatase and osteocalcin among the three groups of the rats. The findings from this study demonstrated that plant protein (soy protein) is beneficial of bone mineral density because it had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein). However, the supplementation of sulfur amino acid on soy results were consistent with prior studies that dietary sulfur amino acid load had a negative effect on calcium balance. The rats fed sulfur amino acid supplementation diet increased urinary calcium excretion and decreased calcium efficiency for total and femur mineral density. Therefore, dietary protein source and sulfur amino acid content influence bone metabolism. (Korean J Nutrition 37(2): 100-107, 2004)

Functional Roles of a Putative B' Delta Regulatory Subunit and a Catalytic Subunit of Protein Phosphatase 2A in the Cereal Pathogen Fusarium graminearum

  • Kim, Hee-Kyoung;Yun, Sung-Hwan
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.259-269
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    • 2012
  • Protein phosphatase 2A (PP2A), a family of serine/threonine protein phosphatases, plays an important role in balancing the phosphorylation status of cellular proteins for regulating diverse biological functions in eukaryotic organisms. Despite intensive studies in mammals, limited information on its role is available in filamentous fungi. Here, we investigated the functional roles of genes for a putative B' delta regulatory subunit (FgPP2AR) and a catalytic subunit (FgPP2AC) of PP2A in a filamentous ascomycete, Fusarium graminearum. Molecular characterization of an insertional mutant of this plant pathogenic fungus allowed us to identify the roles of FgPP2AR. Targeted gene replacement and complementation analyses demonstrated that the deletion of FgPP2AR, which was constitutively expressed in all growth stages, caused drastic changes in hyphal growth, conidia morphology/germination, gene expression for mycotoxin production, sexual development and pathogenicity. In particular, overproduction of aberrant cylindrical-shaped conidia is suggestive of arthroconidial induction in the ${\Delta}FgPP2AR$ strain, which has never been described in F. graminearum. In contrast, the ${\Delta}FgPP2AC$ strain was not significantly different from its wild-type progenitor in conidiation, trichothecene gene expression, and pathogenicity; however, it showed reduced hyphal growth and no perithecial formation. The double-deletion ${\Delta}FgPP2AR;{\Delta}FgPP2AC$ strain had more severe defects than single-deletion strains in all examined phenotypes. Taken together, our results indicate that both the putative regulatory and catalytic subunits of PP2A are involved in various cellular processes for fungal development in F. graminearum.

Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

PtdIns(3,5)P2 5-phosphatase Fig4 Interacts with Kinesin Superfamily 5A (KIF5A) (PI(3,5)P2 5-phosphatase Fig4와 Kinesin superfamily 5A (KIF5A)의 결합)

  • Jang, Won Hee;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.1
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    • pp.14-19
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    • 2014
  • Kinesin-1 consists of two heavy chains (KHCs), also called KIF5s, and two light chains (KLCs) that form a heterotetrameric complex. Here, we demonstrate the binding of a neuronal KHC, KIF5A, to the carboxyl (C)-terminal tail region of Fig4 (also known as Sac3), a phosphatase that removes the 5-phosphate from phosphatidylinositol-3,5-bisphosphate ($PtdIns(3,5)P_2$). Fig4 bound to the C-terminal region of KIF5A but not to other KHCs (KIF5B and KIF5C) and KLC1 in yeast two-hybrid assays. The interaction was further confirmed in a glutathione S-transferase pull-down assay and by co-immunoprecipitation. Anti-KIF5A antibody co-immunoprecipitated Fig4 with KIF5A from mouse brain extracts. These results suggest that kinesin-1 could transport the Fig4-associated protein complex or cargo in cells.

Protein Tyrosine Phosphatase, Receptor Type B (PTPRB) Inhibits Brown Adipocyte Differentiation through Regulation of VEGFR2 Phosphorylation

  • Kim, Ji Soo;Kim, Won Kon;Oh, Kyoung-Jin;Lee, Eun-Woo;Han, Baek Soo;Lee, Sang Chul;Bae, Kwang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.645-650
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    • 2019
  • Brown adipocytes have an important role in the regulation of energy balance through uncoupling protein-1 (UCP-1)-mediated nonshivering thermogenesis. Although brown adipocytes have been highlighted as a new therapeutic target for the treatment of metabolic diseases, such as obesity and type II diabetes in adult humans, the molecular mechanism underlying brown adipogenesis is not fully understood. We recently found that protein tyrosine phosphatase receptor type B (PTPRB) expression dramatically decreased during brown adipogenic differentiation. In this study, we investigated the functional roles of PTPRB and its regulatory mechanism during brown adipocyte differentiation. Ectopic expression of PTPRB led to a reduced brown adipocyte differentiation by suppressing the tyrosine phosphorylation of VEGFR2, whereas a catalytic inactive PTPRB mutant showed no effects on differentiation and phosphorylation. Consistently, the expression of brown adipocyte-related genes, such as UCP-1, $PGC-1{\alpha}$, PRDM16, $PPAR-{\gamma}$, and CIDEA, were significantly inhibited by PTPRB overexpression. Overall, these results suggest that PTPRB functions as a negative regulator of brown adipocyte differentiation through its phosphatase activity-dependent mechanism and may be used as a target protein for the regulation of obesity and type II diabetes.

Constitutively active Ras negatively regulates Erk MAP kinase through induction of MAP kinase phosphatase 3 (MKP3) in NIH3T3 cells

  • Park, Young Jae;Lee, Jong Min;Shin, Soon Young;Kim, Young Ho
    • BMB Reports
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    • v.47 no.12
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    • pp.685-690
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    • 2014
  • The Ras/Raf/MEK/Erk signaling pathway is important for regulation of cell growth, proliferation, differentiation, survival, and apoptosis in response to a variety of extracellular stimuli. Lack of Erk MAPK activation is observed in several cancer cells despite active activation of Ras. However, little is known about the modulation of Erk1/2 activity by active Ras. Here, we show that overexpression of active H-Ras (H-RasG12R) in NIH3T3 fibroblasts impaired FGF2-induced Erk1/2 phosphorylation, as compared to wild-type cells. Northern blot analysis revealed that prolonged expression of active Ras increased MAP kinase phosphatase 3 (MKP3) mRNA expression, a negative regulator of Erk MAPK. Inhibition of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway abrogated active Ras-induced up-regulation of MKP3 expression, leading to the rescue of Erk1/2 phosphorylation. Our results demonstrated that the Ras/Raf/MEK/Erk signaling cascade is negatively regulated by the PI3K/Aktdependent transcriptional activation of the MKP3 gene.

Gene Cloning, Expression, and Characterization of Glucose-1-Phosphatase from Enterobacter cloacae B11

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
    • Fisheries and Aquatic Sciences
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    • v.13 no.1
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    • pp.49-55
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    • 2010
  • A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

Screening of Bioactive Materials from Freshwater Microalgae (담수산 미세조류로부터 생리활성물질의 탐색)

  • Lee, Wan-Seok;Choi, Ae-Ran;Ahn, Chi-Yong;Oh, Hyun-Cheol;Ahn, Jong-Seog;Oh, Hee-Mock
    • ALGAE
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    • v.19 no.3
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    • pp.271-276
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    • 2004
  • One hundred and fifty four micro algal strains, newly isolated from nationwide freshwaters in Korea, were screened for their anticancer, ant diabetic, and antibiotic activities. The micro algal strains were cultured with different nutritional conditions that were divided into 4 groups as follows; a normal Allen medium, nitrogen (N)-limited medium, phosphorus (P)-limited medium, and N and P-limited medium. Algal biomass was extracted with a mixture of acetone:H₂O (1:1, v:v) and the extracts were used for the screening of bioactive materials. Anticancer, ant diabetic, and antibiotic materials were screened by the methods of vaccinia Hl-related protein tyrosine phosphates (VHR DS-PTPase) inhibition, protein tyrosine phosphates 1B (PTP1B) inhibition, and paper disk. The inhibition activity of VHR DS-PTPase was observed in 18 strains, having a maximum 79% inhibition from Anabaena affinis and the inhibition activity of PTP1B was observed in 9 strains, having a maximum 97% from Sphaerocystis schroeteri. Microcystis aeruginosa incubated in an N and P-limited medium showed antibiotic activity in 8 species out of 13 pathogenic bacteria. As a whole, it seemed that the stressed condition such as N and/or P limitation increased the production of bioactive materials in micro algae.

Protein tyrosine phosphatase profiling studies during brown adipogenic differentiation of mouse primary brown preadipocytes

  • Choi, Hye-Ryung;Kim, Won Kon;Park, Anna;Jung, Hyeyun;Han, Baek Soo;Lee, Sang Chul;Bae, Kwang-Hee
    • BMB Reports
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    • v.46 no.11
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    • pp.539-543
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    • 2013
  • There is a correlation between obesity and the amount of brown adipose tissue; however, the molecular mechanism of brown adipogenic differentiation has not been as extensively studied. In this study, we performed a protein tyrosine phosphatase (PTP) profiling analysis during the brown adipogenic differentiation of mouse primary brown preadipocytes. Several PTPs, including PTPRF, PTPRZ, and DUSP12 showing differential expression patterns were identified. In the case of DUSP12, the expression level is dramatically downregulated during brown adipogenesis. The ectopic expression of DUSP12 using a retroviral expression system induces the suppression of adipogenic differentiation, whereas a catalytic inactive DUSP12 mutant showed no effect on differentiation. These results suggest that DUSP12 is involved in brown adipogenic differentiation and may be used as a target protein for the treatment or prevention of obesity by the regulation of brown adipogenic differentiation.

Brazilin Inhibits Activities of Protein Kinase C and Insulin Receptor Serine Kinase in Rat Liver

  • Kim, Seong-Gon;Kim, You-Me;Khil, Lee-Yong;Jeon, Sun-Duck;So, Dhong-Su;Moon, Chang-Hyun;Moon, Chang-Kiu
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.140-146
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    • 1998
  • Hypoglycemic action of brazilin was found to be based on the improvement of peripheral glucose utility, and this action might be correlated with the insulin action pathway. In the present study we investigated the effect of brazilin on the insulin receptor autophosphorylation, protein kinase C (PKC), protein phosphatase and insulin receptor serine kinase in order to confirm whether the hypoglycemic mechanism is concerned with insulin action pathway. Brazilin was found to inhibit PKC and insulin receptor serine kinase, which are involved in the regulation of insulin signal pathway. But any significant effect was not shown on insulin receptor tyrosine kinase activity, autophosphorylation and phosphatase activity. These findings suggest that brazilin might enhance insulin receptor function by decreasing serine phosphorylation, which might mediate hypoglycemic effect of brazilin.

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