• BMB Reports
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• v.29 no.3
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• pp.236-240
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• 1996

A 23-kDa molecular mass of antioxidant protein was purified from human liver. This protein exhibited the preventive effect against the inactivation of glutamine synthetase by a metal-catalyzed oxidation system. This antioxidant activity was supported by a thiol-reducing equivalent such as dithiothreitol in a similar manner to that of the 25-kDa thiol-specific antioxidant protein (TSA) from human red blood cells (HR). However, a thioredoxin-linked peroxidase activity of thiol-specific antioxidant protein of human liver (HLTSA) (0.91 ${\mu}mol/min/nmol$ of HLTSA) was much lower than that of thiol-specific antioxidant protein of human red blood cells (HRTSA) (16.4 ${\mu}mol/min/nmol$ of HRTSA). This HLTSA is also immnologically distinct from HRTSA Amino acid sequences of the three tryptic peptides (P1, P2, P3) of HLTSA were found to be completely homologous to segments of the known Mer5-like protein, which belongs to the known TSA family.

• Genomics & Informatics
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• v.16 no.4
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• pp.23.1-23.5
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• 2018

Virus taxonomy was initially determined by clinical experiments based on phenotype. However, with the development of sequence analysis methods, genotype-based classification was also applied. With the development of genome sequence analysis technology, there is an increasing demand for virus taxonomy to be extended from in vivo and in vitro to in silico. In this study, we verified the consistency of the current International Committee on Taxonomy of Viruses taxonomy using an in silico approach, aiming to identify the specific sequence for each virus. We applied this approach to norovirus in Caliciviridae, which causes 90% of gastroenteritis cases worldwide. First, based on the dogma "protein structure determines its function," we hypothesized that the specific sequence can be identified by the specific structure. Firstly, we extracted the coding region (CDS). Secondly, the CDS protein sequences of each genus were annotated by the conserved domain database (CDD) search. Finally, the conserved domains of each genus in Caliciviridae are classified by RPS-BLAST with CDD. The analysis result is that Caliciviridae has sequences including RNA helicase in common. In case of Norovirus, Calicivirus coat protein C terminal and viral polyprotein N-terminal appears as a specific domain in Caliciviridae. It does not include in the other genera in Caliciviridae. If this method is utilized to detect specific conserved domains, it can be used as classification keywords based on protein functional structure. After determining the specific protein domains, the specific protein domain sequences would be converted to gene sequences. This sequences would be re-used one of viral bio-marks.

• Korean journal of applied entomology
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• v.34 no.2
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• pp.140-146
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• 1995

Changes in protein content during oocyte development was measured and egg-specific protein was characterized from the eggs in Lucilia illustris. During normal development ovarian protein was rapidly increased at 72hr and reached maximum at 96hr after a protein meal, when the eggs were fully matured. Purified protein from the ovaries by gel filtration of DEAE-cellulose an Sephacryl S-200 was loaded on 7.5% native polyacrylamide gel electrophoresis and identified at ${R}_{f}$ 0.4 as egg-specific protein, which has a mol. wt of 110,000. A total of 13 amino acids in th egg-specific protein was identified and expecially asparagine, glutamic acid, and tyrosine were highly concentrated. Five fatty acids were also identified. It is suggested that there is a specific protein in the eggs of L. illustris except yolk protein synthesized and secreted by fat body.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.396-403
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• 1997

In order to direct the form of the immune response in an antigen-specific manner, we constructed a fusion protein (OVA/IL12) that contained the T cell-dependent antigen, ovalbumin (OVA), covalently linked to murine interleukin-12 (IL-12). The OVA/IL12 protein was produced in a baculovirus expression system and was purified by anti-OVA immunoaffinity chromatography. The purified OVA/ILI2 protein displayed potent IL-12 bioactivity in an IL-12 proliferation assay. BALB/c mice immunized with the OVA/IL12 protein produced increased quantities of anti-OVA IgG2a antibody compared with mice immunized with recombinant OVA alone. Lymph node cells from the immunized mice with the OVA/IL12 protein produced large amounts of IFN-,Y when restimulated in vitro with OVA, while those from mice immunized with the OVA protein produced little or no IFN-.gamma.. In contrast, immunization with a mixture of OVA and free recombinant IL-12 also induced IFN-.gamma. production, which was not OVA-specific. These studies indicate that the OVA/IL12 fusion protein can induce OVA-specific, Th1-dominated immune responses, and that the covalent linkage of OVA and IL-12 confines the effect of IL-12 to OVA-specific cells.

• The Journal of Natural Sciences
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• v.4
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• pp.1-10
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• 1991

The expression of the cloned Saccharomyces cerevisiae antioxidant protein gene in E. coil on plasmid was studied. Introduction of the DNA encoding yeast thiol-specific antioxidant protein into expression vector, PKK 223-3, having the tac promoter resulted in the production in E. coil of a Mr 27,000 polypeptide. This protein represented as much as about 1% of the bacterial soluble protein and showed the thiol-specific antioxidant properties. The bacterial protein showed physico-chemical properties indistringuishable from those of the yeast protein.

• Journal of Nutrition and Health
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• v.29 no.6
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• pp.569-577
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• 1996

This study was performed to determine the effect of maternal protein intake on 1) the concentration of immune substances in milk 2) degree of passive immunity to pups via lactation, and 3) specific antibody production to a specific antigen, $\beta$-lactoglobulin(BLG). 4) the effect of passive immunity that pups received from mother during lactation on the production of antibodies when the pups were challenged to the same antigen. Part of the female rats were immunized with BLG before and during pregnancy. The pregnant rats were placed into either 25% or 10% isolated soy protein diet throughout gestation and lactation. After weaning, pups from each group continued to be fed the same diet. At 18 weeks of age, all the pups were challenged with BLG. Total IgA and IgG, lysozyme, BLG-specific IgA and IgG were measured in dam's serum, dam's milk, and pup's serum. Total IgG, and lysozyme in dam's serum and milk were higher in high protein group. Total IgA and IgG in pup's serum remained higher in high protein group from 5 to 18 weeks of age. BLG-specific antibodies were found in the milk and serum of immunized dams, and in serum of pups born to immunized dams but not in the non-immunized group. BLG-specific IgA and IgG were again higher in high protein group and declined with time. The concentration decreased faster in the low proetein group than in the high protein groups. After immunization the pups with LBG, serum BLG-specific antibodies were not differ between rats born to immunized dams and those born to non-immunized dams. Therefore passive immunity rats received via milk as a pup had no effect on the BLG-specific antibody production later in life. This study shows the importance of protein status of mother and strongly support to the endorsement of breast feeding.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.2
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• pp.155-161
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• 2014

Overexpression of amyloid precursor protein with the Swedish mutation causes abnormal hyperphosphorylation of the microtubule-associated protein tau. Hyperphosphorylated isoforms of tau are major components of neurofibrillary tangles, which are histopathological hallmarks of Alzheimer's disease. Protein phosphatase 2A (PP2A), a major tau protein phosphatase, consists of a structural A subunit, catalytic C subunit, and a variety of regulatory B subunits. The B subunits have been reported to modulate function of the PP2A holoenzyme by regulating substrate binding, enzyme activity, and subcellular localization. In the current study, we characterized regulatory B subunit-specific regulation of tau protein phosphorylation. We showed that the PP2A B subunit PPP2R2A mediated dephosphorylation of tau protein at Ser-199, Ser-202/Thr-205, Thr-231, Ser-262, and Ser-422. Down-regulation of PPP2R5D expression decreased tau phosphorylation at Ser-202/Thr-205, Thr-231, and Ser-422, which indicates activation of the tau kinase glycogen synthase kinase 3 beta ($GSK3{\beta}$) by PP2A with PPP2R5D subunit. The level of activating phosphorylation of the $GSK3{\beta}$ kinase Akt at Thr-308 and Ser-473 were both increased by PPP2R5D knockdown. We also characterized B subunit-specific phosphorylation sites in tau using mass spectrometric analysis. Liquid chromatography-mass spectrometry revealed that the phosphorylation status of the tau protein may be affected by PP2A, depending on the specific B subunits. These studies further our understanding of the function of various B subunits in mediating site-specific regulation of tau protein phosphorylation.

• Journal of Plant Biology
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• v.36 no.3
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• pp.301-304
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• 1993

Root nodule specific proteins of Alnus hirsuta were examined. SDS-PAGE pattern of the Alnus root nodule was simpler than that of soybean, showing five nodule specific proteins whose molecular weights were 48, 40, 36, 26 and 19 kD, respectively. Among them, 48 kD protein existed most abundantly and were composed of two subunits whose pI value were 4.0 and 4.3, respectively. The 48 kD protein seemed to be a heme containing protein based on reaction with diaminobenzidine. Although 19 kD protein was present in small amount, it was most similar to leghemoglobin in terms of its molecular weight.

• IMMUNE NETWORK
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• v.11 no.5
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• pp.268-280
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• 2011

Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

• Journal of Microbiology
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• v.39 no.4
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• pp.314-320
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• 2001

Macrophages stimulated by lipopolysaccharide(LPS) from gram negative bacteria undergo activation of a group of immediate early genes including Rantes. The mouse Rantes gene promoter region contains an LPS rsponsive element(LPE) We detected 3 specific bands termed B1, B2 and 3 formed by the interaction of the LPE and proteins found in LPS-stimulated RAW 367.7 cells. An additional band B4 was determined to be an Ap-1 binding protein. The B1 band appears within 1 hour of LPS nuclear extracts from LPS-stimulation, and this protein kinase enhances B1 and formation. The B1 band can be converted to band B2/B3 by adding specific heparin column fraction purified Ser/Thr specific protein phosphatases PP-1 and PP-2A can stimulate the same conversion to about the same extent. Thus, the formation of the LRE sequence binding complex appears to be regulated by Ser/Thr protein kinase and one or more Ser/Thr specific phosphatases. At least four proteins are involved in the trgulation of the LRE-dependent Rants experssion: two binding factors that bind directly to the target sequences. and two factors that control their binding. The future purification and characterization of these binding pro-teins will reveal in detail the mechanism of Rantes gene activation after LPS stimulation.