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Articles by S. Yokoyama
Total Records ( 8 ) for S. Yokoyama
  K. Kita , S. Okada , H. Sekino , K. Imou , S. Yokoyama and T. Amano
  Botryococcus braunii, a green colonial microalga, is an unusually rich renewable source of hydrocarbons. In this study, wet microalgae harvest was thermally pretreated to enhance hydrocarbon recovery using a solvent extraction process. Samples containing a mixture of B. braunii and water were kept below 100  °C for 10 min. The observed hydrocarbon recovery was 97.8% at 90 °C. The extraction results suggest that the energy-intensive concentration and drying processes of the harvest could be eliminated. The proposed thermal pretreatment would revolutionize the conventional downstream processes.
  The objective of the present study was to investigate the effect of dietary phospholipid (PL) level on growth and feed intake of juvenile amberjack (Seriola dumerili) fed non-fishmeal (non-FM) diet containing alternative protein sources; soybean protein isolate, tuna muscle by-product powder and krill meal. Three non-FM diets were prepared to contain three levels (14, 37 and 54gkg−1 dry diet) of PL (soybean lecithin acetone insoluble, 886gkg−1) and growth performance was monitored in a 30-day growth trial by using 2.6g of fish. The results indicated that final body weight, weight gain and feed intake significantly increased with increasing dietary PL level. At the highest dietary PL level (54gkg−1 dry diet), the fish consumed 14.8% and 10.2% as much feed as those fish fed diets containing 14gkg−1 dry diet and 37gkg−1 dry diet PL, respectively. An increasing tendency with increasing dietary PL level on feed efficiency was observed. In conclusion, the present study demonstrated that dietary PL supplementation could increase feed intake, and improve the growth of juvenile S. dumerili fed non-FM diets. Therefore, purified PL might be a good candidate to stimulate the growth of fish through enhancing the feed intake when they are fed diets containing alternative protein sources.
  This study was conducted to examine the effects of dietary ascorbic acid (AsA) and phospholipid (PL) and their interaction on growth, survival, and stress resistance in red sea bream larvae. Twenty-six days old red sea bream were fed nine micro-bound diets supplemented three levels of AsA (0, 800 and 1600 mg kg−1 diet) and PL (0, 20 and 40 g kg−1 diet) for 15 days. Dietary AsA and PL were both significant factors on survival rates. There was also an interaction between dietary AsA and PL on survival rate (P < 0.05). The larvae fed 800 or 1600 mg kg−1 AsA with 40 g kg−1 PL diets showed the highest survival rate, with values similar to those of the live-food supplemented group. Stress resistance against low salinity exposure significantly increased with increased dietary level of AsA and PL. However, significant interaction of AsA and PL was not detected. The larvae fed 1600 mg kg−1 AsA with 40 g kg−1 PL diet showed the highest stress resistance among all diets, but it was not significantly different than that of larvae fed 800 mg kg−1 AsA with 40 g kg−1 PL diet. This study clearly demonstrated that combined use of AsA and PL can improve survival of 26–40 days posthatching red sea bream larvae. Moreover, the present study suggested that 800 mg kg−1 AsA with 40 g kg−1 PL in diet was needed for producing high quality seedling under the stressful conditions.
  R Lu , J Ito , N Iwamoto , T Nishimaki Mogami and S. Yokoyama

Fibroblast growth factor 1 (FGF-1) enhances apolipoprotein E (apoE) expression and apoE-HDL biogenesis in autocrine fashion in astrocytes (Ito, J., Y. Nagayasu, R. Lu, A. Kheirollah, M. Hayashi, and S. Yokoyama. Astrocytes produce and secrete FGF-1, which promotes the production of apoE-HDL in a manner of autocrine action. J. Lipid Res. 2005. 46: 679–686) associated with healing of brain injury (Tada,T., J-i. Ito, M. Asai, and S. Yokoyama. Fibroblast growth factor 1 is produced prior to apolipoprotein E in the astrocytes after cryo-injury of mouse brain. Neurochem. Int. 2004. 45: 23–30). FGF-1 stimulates mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) to increase cholesterol biosynthesis and phosphatidylinositol 3-OH kinase (PI3K)/Akt to enhance apoE-HDL secretion (Ito, J., Y. Nagayasu, K. Okumura-Noji, R. Lu, T. Nishida, Y. Miura, K. Asai, A. Kheirollah, S. Nakaya, and S. Yokoyama. Mechanism for FGF-1 to regulate biogenesis of apoE-HDL in astrocytes. J. Lipid Res. 2007. 48: 2020–2027). We investigated the mechanism for FGF-1 to upregulate apoE transcription. FGF-1 increased apoE and liver X receptor (LXR) mRNAs in rat astrocytes. Increase of LXR mRNA was suppressed by inhibition of the FGF-1 receptor-1 and MEK/ERK but not by inhibition of PI3K/Akt. The increases of apoE mRNA and apoE-HDL secretion were both inhibited by downregulation or inhibition of LXR, while they were partially suppressed by inhibiting cholesterol biosynthesis. We identified the liver X receptor element responsible for activation of the rat apoE promoter by FGF-1 located between –450 and –320 bp, and the direct repeat 4 (DR4) element in this region (–448 to –433 bp) was responsible for the activation. Chromatin immunoprecipitation analysis supported that FGF-1 enhanced association of LXR with the rat apoE promoter. FGF-1 partially activated the apoE promoter even in the presence of an MEK inhibitor that inhibits the FGF-1-mediated enhancement of cholesterol biosynthesis. On the other hand, FGF-1 induced production of 25-hydroxycholesterol by MEK/ERK as an sterol regulatory element-dependent reaction besides cholesterol biosynthesis. We concluded that FGF-1-induced apoE expression in astrocytes depends on LXR being mediated by both LXR expression and an LXR ligand biosynthesis.

  K Takimoto , M Wakiyama and S. Yokoyama

In mammalian cells, microRNAs (miRNAs) are incorporated into miRNA-induced silencing complexes (miRISCs), which regulate protein expression post-transcriptionally through binding to 3'-untranslated regions of target mRNAs. Argonaute2 (Ago2), a key component of the miRISC, recruits GW182, a component of the processing body (GW/P-body), to the target mRNAs. To elucidate the function of GW182 in an miRNA-mediated translational repression, we analyzed Argonaute-binding sites in GW182. We found that human GW182 contains three binding sites for Ago2, within the amino-terminal glycine tryptophan (GW/WG)-repeated region that is characteristic of the GW182 family proteins. We also found that the first and second Ago2-binding site is conserved within the amino-terminal half of TNRC6B, which is a paralog of GW182. Each of the Ago-binding sites is alone sufficient to bind Ago2. Furthermore, we demonstrated that multiple Argonaute proteins were connected via the GW182 protein. A GW182 fragment containing the Ago2-binding region partially relieved let-7-mediated repression of protein synthesis in a mammalian cell-free system. Coincidentally, let-7-directed target mRNA deadenylation was delayed. Together, these results strongly suggested that the interactions of GW182 with Argonautes may induce the formation of large complexes containing miRNA target mRNAs, and may be critical for miRNA-mediated translational repression.

  T Nishida , J. i Ito , Y Nagayasu and S. Yokoyama

Fibroblast growth factor-1 (FGF-1) is released from astrocytes in stress and stimulates MEK/ERK and PI3K/Akt pathways in autocrine fashion to increase synthesis of cholesterol and 25-OH-cholesterol, and to induce transport and secretion of apoE, respectively. FGF-1-induced phosphorylation of Src, and phosphorylation of MEK, ERK and Ark was inhibited by Src inhibitors in rat astrocytes. Src inhibitors also suppressed FGF-1-induced increase of biosynthesis and release of cholesterol and increase of apolipoprotein E (apoE) secretion. The results were reproduced in rat astrocytoma cells transfected by rat apoE and in 3T3-L1 cells. Down-regulation of Src expression reduced FGF-1-induced phosphorylation of the signalling protein and subsequent reactions. Increase by FGF-1 of messages of apoE and HMG-CoA reductase was not influenced by Src inhibitors or by its down-regulation. We conclude that FGF-1 activates Src for activation of MEK/ERK and PI3K/Akt pathways, while Src may not be involved in enhancement of transcription of the cholesterol-related genes.

  K Kodama , H Nakayama , K Sakamoto , S Fukuzawa , T Kigawa , T Yabuki , M Kitabatake , K Takio and S. Yokoyama

A variety of unique codons have been employed to expand the genetic code. The use of the opal (UGA) codon is promising, but insufficient information is available about the UGA suppression approach, which facilitates the incorporation of non-natural amino acids through suppression of the UGA codon. In this study, the UGA codon was used to incorporate 4-iodo-l-phenylalanine into position 32 of the Ras protein in an Escherichia coli cell-free translation system. The undesired incorporation of tryptophan in response to the UGA codon was completely repressed by the addition of indolmycin. The minor amount (3%) of contaminating 4-bromo-l-phenylalanine in the building block 4-iodo-l-phenylalanine led to the significant incorporation of 4-bromo-l-phenylalanine (21%), and this problem was solved by using a purified 4-iodo-l-phenylalanine sample. Optimization of the incubation time was also important, since the undesired incorporation of free phenylalanine increased during the cell-free translation reaction. The 4-iodo-l-phenylalanine residue can be used for the chemoselective modification of proteins. This method will contribute to advancements in protein engineering studies with non-natural amino acid substitutions.

  G Kawai and S. Yokoyama

The late Prof. Tatsuo Miyazawa was an outstanding physical chemist, who established a number of spectroscopic methods to analyse the structures of proteins, peptides and nucleotides, and used them to understand molecular functions. He developed an infrared spectroscopic method to quantitatively analyse the secondary structures, -helices and β-strands, of proteins. He successfully utilized nuclear magnetic resonance (NMR) methods to determine the conformations of peptides and proteins, particularly with respect to the interactions with their target molecules, which served as a solid basis for the wide range of applications of NMR spectroscopy to life science research. For example, he found that physiologically active peptides are randomly flexible in solution, but assume a particular effective conformation upon binding to their functional environments, such as membranes. He also used NMR spectroscopy to quantitatively analyse the conformer equilibrium of nucleotides, and related the dynamic properties of the modified nucleosides naturally-occurring in transfer ribonucleic acids (tRNAs) to their roles in correct codon recognition in protein synthesis. Furthermore, he studied the mechanisms of protein biosynthesis systems, including tRNA and aminoacyl-tRNA synthetases. Inspired by the structural mechanism of amino acid recognition by aminoacyl-tRNA synthetases, as revealed by NMR spectroscopy, he initiated a new research area in which non-natural amino acids are site-specifically incorporated into proteins to achieve novel protein functions (alloprotein technology).

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