• Title/Summary/Keyword: LIM protein

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Fish Meal Replacement with a Mixture of Plant and Animal Protein Sources in Extruded Pellet (EP) Diet for Red Seabream Pagrus major at Low Water Temperature (저수온기 참돔(Pagrus major) EP사료 내 동·식물성단백질 혼합물의 어분 대체)

  • Lim, Jongho;Kim, Min-Gi;Lim, Hyunwoon;Lee, Bong-Joo;Lee, Seunghyung;Hur, Sang-Woo;Kim, Kang-Woong;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.3
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    • pp.350-357
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    • 2021
  • This study aimed to evaluate how fish meal (FM) replacement in diets with a mixture of animal and plant protein sources affect growth performance, feed utilization, hematological parameters and innate immunity of red seabream Pagrus major. A control FM diet was formulated to contain 65% FM (Con). Two other diets were prepared replacing FM in the control diet with a mixture of protein sources (wheat gluten, soy-protein concentrate, tankage meal, and poultry by-product meal) by 30 and 40% (FM30 and FM40, respectively). Total 300 red seabream (body weight, 77.6±0.3g) were distributed to 12 tanks (300 L) in 4 replicates per diet. The fish were fed the diets to apparent satiation for 19 weeks. After the feeding trial, no significant differences could be observed in growth performance, feed utilization, hematological parameters, innate immunity, and survivals among all the dietary treatments. This long-term feeding trial at low water temperature (13.8-17.5℃) indicates that a proper mixture ratio of wheat gluten, soy protein concentrate, tankage meal, and poultry by-product meal can replace FM up to 40% in red seabream diets.

Phosphorylation of a 66 kDa Protein, a Putative Protein Kinase C Substrate, is Related to Chondrogenesis of Chick Embryo Mesenchymes In Vitro

  • Lee, Sun-Ryung;Sonn, Jong-Kyung;Yoo, Byung-Je;Lim, Young-Bin;Kang, Shin-Sung
    • BMB Reports
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    • v.31 no.4
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    • pp.350-354
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    • 1998
  • To understand the role of protein kinase C (PKC) in the regulation of chondrogenesis, we examined proteins which are phosphorylated by PKC. Stage 23/24 chick embryo wing mesenchymes were micromass-cultured to induce chondrogenesis and cell extracts were phosphorylated in a condition that activates PKC. Several proteins including 63 and 66 kDa proteins were phosphorylated. The 66 kDa protein was phosphorylated only in the presence of phorbol 12-myristate 13-acetate (PMA) and phosphatidylserine CPS), and the phosphorylation was almost completely diminished by bisindolylmaleimide, a PKC inhibitor. In addition, partially purified PKC increased the phosphorylation of the 66 kDa protein. Treatment of cultures with lysophosphatidylcholine (LPC) promoted chondrogenesis and phosphorylation of 66 kDa protein, while PMA and thymeleatoxin inhibited both of the two events. Our results suggest that the 66 kDa protein is a putative substrate of PKC, and phosphorylation of the 66 kDa protein, probably by $PKC\alpha$ is required for chondrogenesis.

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Protein Array Fabricated by Microcontact Printing for Miniaturized Immunoassay

  • Lee Woo-Chang;Lim Sang-Soo;Choi Bum-Kyoo;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1216-1221
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    • 2006
  • A protein array was fabricated for a miniaturized immunoassay using microcontact printing ($\mu$CP). A polydimethylsiloxane (PDMS) stamp with a 5 $\mu$m$\times$5 /$\mu$m dimension was molded from a silicon master developed by photolithography. Under optimal fabrication conditions, including the baking, incubation, and exposure time, a silicon master was successfully fabricated with a definite aspect ratio. An antibody fragment was utilized as the ink for the $\mu$CP, and transferred to an Au substrate because of the Au-thiol (-SH) interaction. The immobilization and antibody-antigen interaction were investigated with fluorescence microscopy. When human serum albumin (HSA) was applied to the protein array fabricated with an antibody against HSA, the detection limit was 100 pg/ml of HSA when using a secondary antibody labeled with a fluorescence tag. The fabricated protein array maintained its activity for 14 days.

THE PROTEIN COMPOSITION OF GINGIVAL CREVICULAR FLUID AND SERUM SAMPLED FROM NORMAL SUBJECTS (정상 치은 열구액과 혈청 단백질 조성에 관한 연구)

  • Lim, Jong-Deuk;Moon, Jin-Kyun;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.24 no.1
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    • pp.178-184
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    • 1994
  • This study was undertaken to examine the protein content of GCF and serum from noraml population in order to standardize the sample loading on SDS/PAGE gels. The resulats were as follows ; 1. The protein concentration of serum was not different between normal group and diseased group. 2. In GCF, the bands of lower molecular weight than albumin were heavily stained, but in serum, the protein bands of higher molecalar weight were found. 3. The profile of protein in normal GCF was characterized by heavily staining bands at 77, 66, 55, 26 KDa corresponding to the positions of transferrin, albumin, heavy and light chains of Ig G. Also 47, 37 KDa nonplasma proteins were found.

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Nucleotide and Deduced Amino Acid Sequences of Rat Myosin Binding Protein H (MyBP-H)

  • Jung, Jae-Hoon;Oh, Ji-Hyun;Lee, Kyung-Lim
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.712-717
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    • 1998
  • The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin- binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7kDa and includes the common consensus 1CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III- Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains `RKPS` sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC)phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4N-myristoylation site.

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Development of Methods for Protein Extraction from Three Major Korean Fermented Soy Foods for 2-Dimensional Gel and Mass Spectrometric Analyses

  • Lim, Jin-Kyu
    • Journal of Applied Biological Chemistry
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    • v.51 no.3
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    • pp.88-94
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    • 2008
  • Three different protein extraction methods-phenol extraction, trichloroacetic acid (TCA) precipitation, and desalting/TCA precipitation-were compared to determine the optimal reproducible high resolution 2-dimensional (2-D) electrophoresis for each chungkugjang, doenjang, and kochujang samples. The soluble proteins from Chungkugjang extracted by phenol were separated with high reproducibility and resolution, and gained 1.75- to 3-fold more protein spots on 2-D gel than those from the other methods. On the contrary, the extracted proteins from doenjang and kochujang treated by desalting/TCA precipitation method showed about 1.5- to 3.3-fold more protein spots on 2-D gel. Using the established methods, the changes in the protein profiles of the fermented soy foods were monitored during the fermentation period by 2-DE. One of the major proteins in soy, $\beta$-conglycinin $\alpha$-subuint, and some proteins with unknown functions were localized on 2-D gel as the protease-resistant proteins throughout the fermentation period of doenjang. Changes in the protein profile monitored by the established methods can provide basic information on unfolding the mechanisms of the generation of biofunctional activity in the fermented soy foods.

The effect of nanoemulsified methionine and cysteine on the in vitro expression of casein in bovine mammary epithelial cells

  • Kim, Tae-Il;Kim, Tae-Gyun;Lim, Dong-Hyun;Kim, Sang-Bum;Park, Seong-Min;Lim, Hyun-Joo;Kim, Hyun-Jong;Ki, Kwang-Seok;Kwon, Eung-Gi;Kim, Young-Jun;Mayakrishnan, Vijayakumar
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.257-264
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    • 2019
  • Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

Effects of Dietary Protein and Lipid Levels on Growth, Feed Utilization and Body Composition of Adult Starry Flounder (Platichthys stellatus)

  • Lee Jong Ha;Cho Sung Hwoan;Lim Han Kyu;Kim Kyoung-Duck;Lee Sang-Min
    • Fisheries and Aquatic Sciences
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    • v.7 no.4
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    • pp.184-191
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    • 2004
  • A 25-week feeding trial of two dietary protein (47 and $52\%$) and three dietary lipid level (7, 12 and $17\%$) factorial design with three replications were conducted to determine effects of dietary protein and lipid levels on growth, feed utilization and body composition of adult starry flounder (Platichthys stellatus), average initial weight 332 g, during the winter season. Survival of fish was not affected by either dietary protein or dietary lipid level. Weight gain, feed efficiency and protein efficiency ratio improved with dietary protein and lipid levels except for those of fish fed the $52\%$ protein diet with $17\%$ lipid. The best growth and feed utilization were observed in the $52\%$ protein diet with $12\%$ lipid, but were not significantly different from those of fish fed the $52\%$ protein diet with $17\%$ lipid or the $47\%$ protein diets with $17\%$ lipid levels. Hepatosomatic and visceral somatic indexes were significantly influenced by dietary protein level, but not by dietary lipid level. None of moisture, crude protein, crude lipid, or glycogen contents of dorsal muscle or liver in starry flounder except for crude lipid in dorsal muscle was significantly influenced by either dietary protein or dietary lipid level. Plasma cholesterol concentration was significantly influenced by both dietary protein and dietary lipid levels. The results of this study suggest that the diets containing $47\%$ protein with $17\%$ lipid or $52\%$ protein with $12-17\%$ lipid are optimal for growth and feed utilization of adult starry flounder under these experimental conditions.

Effect of Chongmyung-Tang Prescription Combination on the Production of Amyloid β protein and β-site amyloid precursor protein-cleaving enzyme Activity in vitro (In vitro에서 β-site amyloid precursor protein-cleaving enzyme 활성과 amyloid β protein 생산에 대한 총명탕가미방(聰明湯加味方)의 효과)

  • Lim, Jung-Hwa;Jung, In-Chul;Lim, Jong-Soon;Kim, Seung-Hyung;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.191-200
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    • 2010
  • Objectives : This experiment was designed to investigate the effect of Chongmyung-Tang Prescription Combination(CmTP-$C_{1-10}$) extract on the production of amyloid $\beta$ protein and $\beta$-site amyloid precursor protein-cleaving enzyme(BACE) activity. Methods : The effect of CmTP-$C_{1-10}$ extract on expression of APP mRNA, BACE2 mRNA in BV2 microglia cell line treated by lipopolysacchride(LPS) and amyloid $\beta$ protein fragment(A$\beta$ fragment) were investigated. The effect of CmTP-$C_{1-10}$ extract on production of amyloid $\beta$ protein(A$\beta$) in BV2 microglia cell line treated by LPS and A$\beta$ fragment were investigated. The effect of CmTP-$C_{1-10}$ extract on BACE activity were investigated. Results : 1. CmTP-$C_9$ extract the most significantly suppressed the expression of APP mRNA, BACE2 mRNA in BV2 microglia cell line treated by LPS and A$\beta$ fragment. 2. CmTP-$C_9$ extract significantly suppressed the production of A$\beta$ in BV2 microglia cell line treated by LPS and A$\beta$ fragment. 3. CmTP-$C_9$ extract the most significantly inhibited BACE activity. Conclusions : These results suggest that CmTP-$C_9$ may be effective for the prevention and treatment of Alzheimer's Disease. Investigation into clinical use of CmTP-$C_9$ for Alzheimer's Disease is suggested for future research.