• Journal of Microbiology and Biotechnology
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• v.2 no.2
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• pp.122-128
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• 1992

Post-translational modifications of the nucleocapsid protein of bovine coronavirus (Quebec strain) were investigated. Coronavirions were radiolabelled in vivo with inorganic $[^{32}P]$orthophosphate and analysed by SDS-PAGE, followed by autoradiography. A single polypeptide with a migration rate of 55 KDa was identified by metabolic phosphate labelling, demonstrating that the nucleocapsid protein of bovine coronavirus was a phosphoprotein. A gene encoding the nucleocapsid protein was inserted immediately downstream from the polyhedrin promoter of Autographa californica nuclear polyhedrosis baculovirus. Spodoptera frugiperda cells infected with this recombinant baculovirus synthesized a 55 KDa polypeptide, as demonstrated by immunoprecipitation with anti-nucleocapsid monoclonal antibody. The recombinant nucleocapsid protein synthesized in Spodoptera cells could also be labelled by $[^{32}P]$orthophosphate. Phosphoamino acid analysis showed that both serine and threonine residues were phosphorylated in authentic, as well as in recombinant nucleocapsid proteins, with a relative phosphorylation ratio of 7:3. Our studies demonstrated that the nucleocapsid protein of bovine coronavirus was a serine and threonine-phosphorylated protein and that Spodoptera insect cells were able to properly phosphorylate the relevant foreign proteins.

• BMB Reports
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• v.32 no.5
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• pp.468-473
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• 1999

Nuclear Factor 1 (NF1) proteins are a family of transcriptional factors consisting of four different types: NF1-A, -B, -C, and -X. Some NF1 transcription factors contain a heptasequence motif, SPTSPSY, which is found as a repeat sequence in the carboxy terminal domain (CTD) of the largest subunit of RNA polymerase II. A similar heptasequence, SPTSPTY, is contained in rat liver NF1-A at a position between residues 469 and 475. In order to investigate the roles of the individual amino acids of the heptasequence of rat liver NF1-A in transcriptional activation, we systematically substituted single and multiple amino acid residues with alanine residue(s) and evaluated the transcriptional activities of the mutated NF1-A. Substitution of a single amino acid reduced transcriptional activity by 10 to 30%, except for the proline residue at position 473, whose substitution with alanine did not affect transcriptional activity. However, changes of all four serine and threonine residues to alanine or of the tyrosine residue along with the serine residue at position 469 to alanine reduced the activity to almost background levels. Our results indicate that multiple serine and threonine residues, rather than a single residue, may be involved in the modulation of the transcriptional activities of the factor. Involvement of the tyrosine residue is also implicated.

• The Korean Journal of Zoology
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• v.4 no.2
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• pp.13-19
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• 1961

Free amino acids at five different developemntal stages (Gastrulation-Hatching -out stage) of Hynobius leechi BOULENGER were analyzed qualitatively by the use of paper paitition chromatography. It was found that the number of free amino acids increased as the development proceeded. The free amino acids identified at each stages are as follows : Gastrulation stage : Alaninie, Aspartic acid, Glutamin acid, Histidine, Methionine. Neural plate formation stage : Alanine , Aspartic acid, Glutamic acid, Glycine, Histidine, MEthionine, Phenylalanine, Proline, Serine, Trypotophan. Middle tail-bud stage : Alanine, Arginine, Asparagine,Aspartic acid, Citrulline, Glutamic acid, Glycine, Histidie,Hydroxyproline, Proline, Leucine, Methionine, Ornithine, Phenylalanine, Serine, Threonine, Tryptophan. Late tail-bud stage : Alanine, Arginine, Asparagine, Aspartic acid, Citrulline, Glutamic acid. Glycine, Histidine, Hydroxyproline, Leucine, Methionine, Ornithine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Valine. Hatching -out stage : the same with the late tail-bud stage. It seems probable that the metabolic systems of amino acids before and after the middle tail-bud stage are quite different from each other. Before the middle tail=-bud stage, the reaction system of amino acids is thought not to be completed while after that stage the system has been completed , because in the former period of the development , the number of freeamino acids increased rapidly with the development , and after that stage, there is practically no change in the number of free amino acids.

• Molecules and Cells
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• v.44 no.7
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• pp.444-457
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• 2021

Although the mechanism of chronic myeloid leukemia (CML) initiation through BCR/ABL oncogene has been well characterized, CML cell differentiation into erythroid lineage cells remains poorly understood. Using CRISPR-Cas9 screening, we identify Chromobox 8 (CBX8) as a negative regulator of K562 cell differentiation into erythrocytes. CBX8 is degraded via proteasomal pathway during K562 cell differentiation, which activates the expression of erythroid differentiation-related genes that are repressed by CBX8 in the complex of PRC1. During the differentiation process, the serine/threonine-protein kinase PIM1 phosphorylates serine 196 on CBX8, which contributes to CBX8 reduction. When CD235A expression levels are analyzed, the result reveals that the knockdown of PIM1 inhibits K562 cell differentiation. We also identify TRIM28 as another interaction partner of CBX8 by proteomic analysis. Intriguingly, TRIM28 maintains protein stability of CBX8 and TRIM28 loss significantly induces proteasomal degradation of CBX8, resulting in an acceleration of erythroid differentiation. Here, we demonstrate the involvement of the CBX8-TRIM28 axis during CML cell differentiation, suggesting that CBX8 and TRIM28 are promising novel targets for CML research.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1741-1747
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• 2012

Water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface of the nanocrystals with conventional aminoacids ligands: serine and threonine. The aminoacids capped ZnS:Mn nanocrystal powders were characterized by XRD, HR-TEM, EDXS, ICP-AES and FT-IR spectroscopy. The optical properties were also measured by UV/Vis and solution photoluminescence (PL) spectroscopies in aqueous solvents. The solution PL spectra showed broad emission peaks around 600 nm with PL efficiencies of 9.7% (ZnS:Mn-Ser) and 15.4% (ZnS:Mn-Thr) respectively. The measured particle sizes for the aminoacid capped ZnS:Mn nanocrystals by HR-TEM images were about 3.0-4.0 nm, which were also supported by Debye-Scherrer calculations. In addition, cytotoxic effects of four aminoacids capped ZnS:Mn nanocrsystals over the growth of wild type E. coli were investigated. Although toxicity in the form of growth inhibition was observed with all the aminoacids capped ZnS:Mn nanocrystals at higher dose (1 mg/mL), ZnS:Mn-Met and ZnS:Mn-Thr appeared non-toxic at doses less than 100 ${\mu}g$/mL. Low biological toxicities were seen at doses less than 10 ${\mu}g$/ mL for all nanocrystals.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.3
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• pp.293-303
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• 1999

The influences of specific protein phosphatase and protein kinase inhibitors on the ATP-sensitive $K^+\;(K_{ATP})$ channel-opening effect of pinacidil were investigated in single rat ventricular myocytes using patch clamp technique. In cell-attached patches, pinacidil $(100\;{\mu}M)$ induced the opening of the $K_{ATP}$ channel, which was blocked by the pretreatment with H-7 $(100\;{\mu}M)$ whereas enhanced by the pretreatment with genistein $(30\;{\mu}M)$ or tyrphostin A23 $(10\;{\mu}M)$. In inside-out patches, pinacidil $(10\;{\mu}M)$ activated the $K_{ATP}$ channels in the presence of ATP (0.3 mM) or AMP-PNP (0.3 mM) and in a partial rundown state. The effect of pinacidil $(10\;{\mu}M)$ was not affected by the pretreatment with protein tyrosine phosphatase 1B $(PTP1B,\;10\;{\mu}g\;ml^{-1}),$ but blocked by the pretreatment of protein phosphatase 2A $(PP2A,\;1\;U\;ml^{-1})$. In addition, pinacidil $(10\;{\mu}M)$ could not induce the opening of the reactivated $K_{ATP}$ channels in the presence of H-7 $(100\;{\mu}M)$ but enhanced it in the presence of ATP (1 mM) and genistein $(30\;{\mu}M).$ These results indicate that the $K_{ATP}$ channel-opening effect of pinacidil is not mediated via phosphorylation of $K_{ATP}$ channel protein or associated protein, although it still requires the phosphorylation of serine/threonine residues as a prerequisite condition.

• BMB Reports
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• v.40 no.5
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• pp.749-756
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• 2007

Phosphorylation on serine/threonine or tyrosine residues of target proteins is an essential and significant regulatory mechanism in signal transduction during many cellular and life processes, including spermatogenesis, oogenesis and fertilization. In the present work, we reported the isolation and characterization of mouse testis-specific serine/threonine kinase 5 (Tssk5), which contains four alternatively spliced variants including, Tssk5$\alpha$, Tssk5$\beta$, Tssk5$\gamma$ and Tssk5$\delta$. Moreover, the locus of Tssk5 is on chromosome 14qC3 and the four variants had a similar high expression in the testis and the heart; however, had a low expression in other tissues, except for Tssk5$\alpha$ which also had comparably high expression in the spleen. Each variant of Tssk5 expression began in the testis 16 days after birth. Aside from TSSK5$\alpha$, the other isoforms have an insertion of ten amino acid residues (RLTPSLSAAG) in region VIb (HRD domain) (His-Arg-Asp). Moreover, only TSSK5$\alpha$ exhibited kinase activity and consistently, a further Luciferase Reporter Assay demonstrated that TSSK5$\beta$, TSSK5$\gamma$ and TSSK5$\delta$ cannot be stimulated at the CREB/CRE responsive pathway in comparison to TSSK5$\alpha$. These findings suggest that TSSK5$\beta$, TSSK5$\gamma$, TSSK5$\delta$ may be pseudokinases due to the insertion, which may damage the structure responsible for active kinase activity. Pull-down assay experiments indicated that TSSK5$\beta$, TSSK5 $\gamma$ and TSSK5$\delta$ can directly interact with TSSK5$\alpha$. In summary, these four isoforms with similar expression patterns may be involved in spermatogenesis through a coordinative way in testis.

• Journal of the Korean Chemical Society
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• v.40 no.8
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• pp.542-548
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• 1996

Electrodes modified with threonine, methionine and serine as ligands, which are incorporated by ion exchange into a polycationic film of electropolymerized $[Ru(v-bpy)_3]^{2+}$, have been employed in the determination of mercury in solution. The redox response of the surface-immobilized mercury/ligand complex was used as the analytical signal. When the polymeric film was electropolymerized, the supporting electrolytes were TBAP and $KPF_6$ to compare the morphology and anodic stripping of resulted polymer electrodes. At the case of the latter, the film had high porosity to give an easy incorporation of dopant anions into polymeric film matrix and a high sensitivity in determination of mercury ion. Especially, this polymer modified electrode exhibited possibility of multiple use in mercury determination over ten times. In all cases, calibration curves which were plotted by log of the surface coverage-normalized redox response vs. log[Hg] exhibited an excellent correlation (r=0.99) for mercury concentrations ranging from 1.0{\times}10^{-8}{\sim}1.0{\times}10^{-2}M$. At these curves relative standard deviation was 5∼8% and saturation response was not observed at high concentration region. Serine of the employed ligands had the best sensitivity in analytical application, which had greater stability constant in forming a complex with mercury than others as $pK_{Hg}=8.54$. The formation constants of threonine and methionine were respectively 7.04 and 7.80.

• Korean Journal of Food Science and Technology
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• v.5 no.2
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• pp.71-77
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• 1973

The results, which was analytically surveyed the free amino acids by the automatic amino acid analyzer adding the enzyme on the Korean cow's fore shank muscles, are as follows: 1. The content of free amino acids in the fore shank muscles, without addition of the enzyme orderly contains alanine, glutamic acid, lysine, glycine, histidine, leucine, threonine, arginine, cystine, serine, proline, isoleucine, phenylalanine, tyrosine, methionine, aspartic acid and valine. 2. In accordance with the addition of the enzyme, by 0.01%, 0.05% and 0.1% the nine free amino acids of glutamic acid, glycine, alanine, cystine, valine, isoleucine, leucine, lysine and arginine were continuosly increased. 3. Proline and histidine were decreased at the enzyme addition of 0.01% after showing the high content at the control, but the quantity of free amino acids was increased according to the increase of the quantity of the enzyme. 4. Aspartic acid, threonine, serine, methionine, tyrosine and phenylalanine were increased till the enzyme addition of 0.05% and remarkably decreased from 0.1%. 5. At cooking the meat, the quantity of the enzyme addition was most effective at 0.05% of meat weight.

• Korean Journal of Breeding Science
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• v.40 no.3
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• pp.269-277
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• 2008

Amino acid composition in grain of Korean native rice core collections was evaluated to identify the useful genotypes for quality enhancing rice breeding and value-added food development. Large variations were found in the contents of 16 amino acids among 388 genotypes tested with 14.9% of the minimum coefficient of variation (CV), and the contents of methionine and histidine showed the highest CV and the second high, respectively. The average values of total amino acid, total essential amino acid, lysine and threonine contents of Korean native rice collections were almost similar with those of cultivating japonica rice, Saechucheong. Among 388 native core collections Baekkokna, Daesona and Saducho were identified as high amino acid germplasm in total amino acid, total essential amino acid, lysine and threonine contents. And Seoksanjo and Sodujo were identified as low amino acid collections. Rice collections showing purple apiculus color tended to distribute in lower serine, proline, glutamic acid, glycine, alanine, valine, leucine, phenylalanine, arginine, and total amino acid contents compared with those of yellowish apiculus color collections. The glutinous rice collections showed the tendency of higher contents of serine, glutamic acid, alanine, valine and leucine compared with those of non-glutinous collections.