• Title/Summary/Keyword: Aldolase A

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Molecular Cloning and Characterization of Salt-inducible Aldolase from Salicornia herbacea (퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석)

  • Cha, Joon-Yung;Netty Ermawati;Kim, Soon-Gil;Lee, Jeung-Joo;Lim, Chae-Oh;Chung, Woo-Sik;Lee, Kon-Ho;Son, Dae-Young
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.323-328
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    • 2003
  • Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

Glyceraldehyde-3-Phosphate, a Glycolytic Intermediate, Plays a Key Role in Controlling Cell Fate Via Inhibition of Caspase Activity

  • Jang, Mi;Kang, Hyo Jin;Lee, Sun Young;Chung, Sang J.;Kang, Sunghyun;Chi, Seung Wook;Cho, Sayeon;Lee, Sang Chul;Lee, Chong-Kil;Park, Byoung Chul;Bae, Kwang-Hee;Park, Sung Goo
    • Molecules and Cells
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    • v.28 no.6
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    • pp.559-563
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    • 2009
  • Glyceraldehyde-3-phosphate is a key intermediate in several central metabolic pathways of all organisms. Aldolase and glyceraldehyde-3-phosphate dehydrogenase are involved in the production or elimination of glyceraldehyde-3-phosphate during glycolysis or gluconeogenesis, and are differentially expressed under various physiological conditions, including cancer, hypoxia, and apoptosis. In this study, we examine the effects of glyceraldehyde-3-phosphate on cell survival and apoptosis. Overexpression of aldolase protected cells against apoptosis, and addition of glyceraldehyde-3-phosphate to cells delayed apoptosis. Additionally, delayed apoptotic phenomena were observed when glyceraldehyde-3-phosphate was added to a cell-free system, in which artificial apoptotic process was induced by adding dATP and cytochrome c. Surprisingly, glyceraldehyde-3-phosphate directly suppressed caspase-3 activity in a reversible noncompetitive mode, preventing caspase-dependent proteolysis. Based on these results, we suggest that glyceraldehyde-3-phosphate, a key molecule in several central metabolic pathways, functions as a molecule switch between cell survival and apoptosis.

Expression and Characterization of a Novel Deoxyribose 5-Phosphate Aldolase from Paenibacillus sp. EA001

  • Kim, Yong-Mo;Choi, Nack-Shick;Kim, Yong-Ook;Son, Dong-Ho;Chang, Young-Hyo;Song, Jae-Jun;Kim, Joong-Su
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.995-1000
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    • 2010
  • A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding a protein of 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79% identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into an expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified using Ni-NTA affinity chromatography and then characterized. The optimum temperature and pH of the enzyme were $50^{\circ}C$ and 6.0, respectively. The specific activity for the substrate deoxyribose 5-phosphate (DR5P) was $62\;{\mu}mol/min/mg$. The $K_m$ value for DR5P was determined to be 145 mM with the $k_{cat}$ value of $3.2{\times}10^2/s$ from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).

Protective and Anti-Pathology Effects of Sm Fructose-1,6-Bisphosphate Aldolase-Based DNA Vaccine against Schistosoma mansoni by Changing Route of Injection

  • Saber, Mohamed;Diab, Tarek;Hammam, Olft;Karim, Amr;Medhat, Amina;Khela, Mamdouh;El-Dabaa, Ehab
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.155-163
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    • 2013
  • This study aimed to evaluate the efficacy of fructose-1,6-bis phosphate aldolase (SMALDO) DNA vaccination against Schistosoma mansoni infection using different routes of injection. The SMALDO has been cloned into the eukaryotic expression vector pcDNA3.1/V5-His TOPO-TA and was used in injecting Swiss albino mice intramuscularly (IM), subcutaneously (SC), or intraperitoneally (IP) ($50{\mu}g/mouse$). Mice vaccinated with non-recombinant pcDNA3.1 served as controls. Each group was immunized 4 times at weeks 0, 2, 4, and 6. Two weeks after the last booster dose, all mice groups were infected with 80 S. mansoni cercariae via tail immersion. At week 8 post-infection, animals were sacrificed for assessment of parasitological and histopathological parameters. High anti-SMALDO IgG antibody titers were detected in sera of all vaccinated groups (P<0.01) compared to the control group. Both the IP and SC vaccination routes resulted in a significant reduction in worm burden (46.2% and 28.9%, respectively, P<0.01). This was accompanied by a significant reduction in hepatic and intestinal egg counts (41.7% and 40.2%, respectively, P<0.01) in the IP group only. The number of dead eggs was significantly increased in both IP and IM groups (P<0.01). IP vaccination recorded the highest significant reduction in granuloma number and diameter (54.7% and 29.2%, respectively, P<0.01) and significant increase in dead miracidia (P<0.01). In conclusion, changing the injection route of SMALDO DNA vaccination significantly influenced the efficacy of vaccination. SMALDO DNA vaccination via IP route could be a promising protective and antipathology vaccine candidate against S. mansoni infection.

Crystallographic snapshots of active site metal shift in E. coli fructose 1,6-bisphosphate aldolase

  • Tran, Huyen-Thi;Lee, Seon-Hwa;Ho, Thien-Hoang;Hong, Seung-Hye;Huynh, Kim-Hung;Ahn, Yeh-Jin;Oh, Deok-Kun;Kang, Lin-Woo
    • BMB Reports
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    • v.49 no.12
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    • pp.681-686
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    • 2016
  • Fructose 1,6-bisphosphate aldolase (FBA) is important for both glycolysis and gluconeogenesis in life. Class II (zinc dependent) FBA is an attractive target for the development of antibiotics against protozoa, bacteria, and fungi, and is also widely used to produce various high-value stereoisomers in the chemical and pharmaceutical industry. In this study, the crystal structures of class II Escherichia coli FBA (EcFBA) were determined from four different crystals, with resolutions between $1.8{\AA}$ and $2.0{\AA}$. Native EcFBA structures showed two separate sites of Zn1 (interior position) and Zn2 (active site surface position) for $Zn^{2+}$ ion. Citrate and TRIS bound EcFBA structures showed $Zn^{2+}$ position exclusively at Zn2. Crystallographic snapshots of EcFBA structures with and without ligand binding proposed the rationale of metal shift at the active site, which might be a hidden mechanism to keep the trace metal cofactor $Zn^{2+}$ within EcFBA without losing it.

Effect of Honey on the Change of Enzyme Activity in Rats (벌꿀이 흰쥐의 효소활성에 미치는 영향)

  • 정동현;백승화;박성수
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.65-73
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    • 1997
  • The purpose of the study was to find an effect of honey on the enzyme activity of Sprague Dawley rats. All experimental rats were fed ad libitum for seven weeks with 68% saccharide diet and at same time fed administratively with 10% and 20% water solution of acacia, sumac, polyflower honey, and sucrose, respectively. The level of LDH activity in serum of rat taken diet with acasia, sumac, and polyflower honey were increased in comparison with the control group. The level of $\alpha$-HBDH activity in serum of rat taken diet with acasia, sumac, polyflower honey, and sugar solution were increased than that other honey solution. The level of GOT and GPT activity in serum was increased by the feeding of solution of 20% acacia honey. The level of ICD activity in serum of rat taken diet with sumac honey was increased but was decreased notably by the feeding of polyflower honey. The level of G-6-P DH activity in whole blood of rat taken diet with honey solutions were decreased, but the level of aldolase activity in serum of rat taken diet with honey solutions were increased.

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Regulation of Protein Expression in Mouse Liver by Inorganic Arsenic: Proteomic Analysis (무기비소에 의한 마우스 간의 단백질 발현 조절 : 단백체 분석)

  • Jin Bo-Hwan;Seong Je-Kyung;Ryu Doug-Young
    • Environmental Mutagens and Carcinogens
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    • v.26 no.2
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    • pp.35-40
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    • 2006
  • Background: Inorganic arsenic is a human carcinogen that can target the liver, but its carcinogenic mechanisms are still unknown. Inorganic arsenic induces a spectrum of tumors including hepatocellular carcinoma in mice. Methods: Pregnant C3H mice were supplied with drinking water containing 50 ppm sodium arsenite during their pregnancy. The protein expression profile in the liver of 0.5-day-old. male offsprings exposed transplacentally to sodium arsenite was analyzed using protein 2D gel electrophoresis followed by mass spectrometry (MALDI-TOF). Results: Expression of proteins such as hydroxymethylglutaryl-CoA synthase mitochondrial precursor (HMG-CoA synthase), ${\beta}$-actin (cytoplasmic 1) and apolipoprotein A-IV precursor (Apo-AIV) were induced in mouse liver by sodium arsenite, while uricase (urate oxidase), guanine nucleotidebinding protein beta subunit 2-like 1 (RACK1) and fructose-bisphosphate aldolase B (Aldolase 2) were down-regulated. Summary: Expression of proteins that have been implicated in carcinogenesis, such as HMG-CoA, ${\beta}$-actin, and RACK1, was regulated in the liver of mice transplacentally exposed to inorganic arsenic.

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Gene Expression in the Muscles of young and Mature Channel Catfish (Ictalurus punctatus) as Analyzed by Expressed Sequence Tags and Gene Filters

  • Soon-Hag Kim
    • Journal of Aquaculture
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    • v.16 no.1
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    • pp.8-14
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    • 2003
  • To generate expressed sequence tags for genomics research involving genetic linkage analysis, to examine gene expression profiles in muscles of channel catfish in a non-normalized muscle cDNA library, to compare gene expression in young and mature channel catfish muscles using the EST reagents and gene filters to demonstrate the feasibility of functional genomics research in small laboratories. 102 randomly picked cDNA clones were analyzed from the catfish muscle cDNA library. Of the sequences generated, 90.2% of ESTs was identified as known genes by identity comparisons. These 92 clones of known gene products represent transcriptional products of 24 genes. The 10 clones of unknown gene products represent 8 genes. The major transcripts (70.1% of the analyzed ESTs) in the catfish muscle are from many major genes involved in muscle contraction, relaxation, energy metabolism and calcium binding such as alpha actin, creatine kinase, parvalbumin, myosin, troponins, and tropomyosins. Gene expression of the unique ESTs was comparatively studied in the young and adult catfish muscles. Significant differences were observed for aldolase, myostatin, myosin light chain, parvalbumin, and an unknown gene. While myosin light chain and an unknown gene (CM 192) are down-regulated in the mature fish muscle, the aldolase, myostatin, and parvalbumin are significantly up-regulated in the mature fish muscle. Although the physiological significance of the changes in expression levels needs to be further addressed, this research demonstrates the feasibility and power of functional genomics in channel catfish. Channel catfish muscle gene expression profiles provide a valuable molecular muscle physiology blueprint for functional comparative genomics.

Synthesis of L-threo-3,4-Dihydroxyphenylserine(L-threo-DOPS) with Thermostabilized Low-Specific L-Threonine Aldolase from Streptomyces coelicolor A3(2)

  • Baik, Sang-Ho;Yoshioka, Hideki;Yukawa, Hideaki;Harayama, Shigeaki
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.721-727
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    • 2007
  • Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

Effects of Different Music Genres on the Stress Levels of Jeju Crossbred Horses (마방에서 음악장르에 따른 Jeju crossbred의 스트레스 변화 연구)

  • Yoonjeong Jang;Jae-Young Choi;Jongan Lee;Yongjun Kang;Nayoung Kim;Minjung Yoon;Moon-Cheol Shin;Sang-Min Shin;Sangsoo Sun;Jiwoong Lee
    • Journal of Life Science
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    • v.33 no.12
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    • pp.995-1001
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    • 2023
  • This study investigated the effects of three music genres (classical, new age, and rock) on the stress levels of six Jeju crossbred horses (Jeju horse × Thoroughbred) in a horse stable. The horses were exposed to the three genres for seven days, and their stress levels were measured by analyzing physiological markers, including neurotransmitter (cortisol, β-endorphin, dopamine, serotonin, and oxytocin) plasma levels and creatine phosphokinase (CPK) and aldolase serum levels. The neurotransmitter analysis showed significant differences in cortisol levels between classical and new age music exposure. Dopamine levels decreased significantly only with new age exposure. Although there were no significant differences in β-endorphin levels between the three genres, β-endorphin levels decreased with increasing classical and new age music playback times and increased with increasing rock music playback times. There were no significant differences in serotonin levels between the three genres. Oxytocin levels decreased significantly with exposure to classical and rock music. The CPK and aldolase analyses showed that CPK levels decreased significantly only with exposure to new age music and increased after playback ended, while aldolase levels decreased significantly with classical and new age music exposure and increased after playback ended. These findings suggest that classical music and new age are the optimal music genres for the psychological stability of Jeju crossbred horses. Playing back an appropriate music genre could be used to improve breeding and promote the welfare of Jeju crossbred horses.