• BMB Reports
• /
• v.39 no.6
• /
• pp.703-708
• /
• 2006

During infection, the common respiratory tract pathogen Streptococcus pneumoniae encounters several environmental conditions, such as upper respiratory tract, lung tissue, and blood stream, etc. In this study, we examined the effects of blood on S. pneumoniae protein expression using a combination of highly sensitive 2-dimensional electrophoresis (DE) and MALDI-TOF MS and/or LC/ESI-MS/MS. A comparison of expression profiles between the growth in THY medium and THY supplemented with blood allowed us to identify 7 spots, which increased or decreased two times or more compared with the control group: tyrosyl-tRNA synthetase, lactate oxidase, glutamyl-aminopeptidase, L-lactate dehydrogenase, cysteine synthase, ribose-phosphate pyrophosphokinase, and orotate phosphoribosyltransferase. This global approach can provide a better understanding of S. pneumoniae adaptation to its human host and a clue for its pathogenicity.

• Animal cells and systems
• /
• v.4 no.2
• /
• pp.145-150
• /
• 2000

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

• The Korean Journal of Zoology
• /
• v.35 no.3
• /
• pp.322-331
• /
• 1992

To investigate the svnthyesis of muscle proteins during differentiation of chicken myoblast, cvtosolic and membrane fractions were used for both sodium dodecvl sulfate polvcrylamide gel eBectrophoresis and two-dimensional gel electrophoresis. An extensive cell fusion was observed in 4 day culture. In the protein pattern of the cvtosolic fraction from SDS-PAGE. several protein bands including 250 kDa and 46 kDa showed remarkable changes during culture. the protein of 46 kDa was the most prominent one ann its optical density was the highest in 5 day culture (OD = 1.30). In the membrane fraction, band of 19.8 kDa showed the highest absorbance with 0.93 OD at 12 hr after initial plating and decreased gradually thereafter to 0.23 in 5 nay culture. From the results of two-dimensional gel electrophoresis of cytosolic fraction, the 46 kDa spot was observed as ko separated forms from culture 2 nary culture, and the sixte of this spot was the largest in 5 nay culture. In the pattern of membrane protein, the extensive appearance of newiv synthesized Proteins was found in a naut culture, but no Prominent spot was observed throughout culture. From the results of the present clay, we found that, during myoblast differentiation, the most prominent proteins were bands of 46 kDa and 19.8 kDa in cvtosolic and membrane fraction, respectively, and the appearance of new proteins was initiated at 48 hr after initial plating, and the 46 kDa protein was predominant in the cytoplasm of late culture in which extensive cell fusion was observed.

• The Journal of Internal Korean Medicine
• /
• v.29 no.2
• /
• pp.469-489
• /
• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

• Journal of Veterinary Clinics
• /
• v.19 no.1
• /
• pp.49-54
• /
• 2002

Korean safflower (Carthami Flos) seed has been known to have healing effects on both bone fracture and osteoporosis. On the base of such a notice, this experiment was carried out to explore the effects of safflower seed on bone formation and bone repair. In addition, the healing mechanism was evaluated by analysing serum after feeding the seed to experimental. animals. The effect of Korean safflower seed were evaluated with 40 rats,3-month old. Forty Sprague-Dawley rats composed of 20 male and 20 female were underwent unilateral tibial defect and then fastened with unilateral fixators. The operated rats were divided into two groups depending on the composition of diet, such as positive control group fed normal diet (C-OP group) and safflower seed group fed 30% of safflower seed diet and 70% of normal diet (S-OP group). Postoperative radiographys were taken once in 2 weeks to evaluate callus formation for operated groups. In addition, a possible protein spots involved in bone recovery were examined using 2-Dimensional Gel Electrophoresis (2-DE). The comparison of the radiography between C-OP and S-OP group were showed that the safflower seed diet appeared to stimulate the formation of callus in the rat. On the images of 2-BE, it was able to identify possible five protein spots, having pl from 4 to 5 and molecular weight range from 24 to 26 kDa, involved in bone formation and repair, since no differing protein spots were found the two between groups except the five spots. No differences were observed between two groups before operation, but clear and bigger protein spots were observed from the S-OP group compared with C-OP group on 6 and 9 weeks post operation. These protein spots were, however, showed similar sizes and densities between two groups in 12 weeks later. The transformation of protein spots was suggested that these protein spots were involved in bone formation and recovery, in addition safflower seed might induce the formation of factors and activate these factors. In conclusion, this study suggest that safflower seed influence a variety of factors in the course of bone formation or the periods of remedy.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.3
• /
• pp.558-562
• /
• 2010

SDS-PAGE of extracted surface-associated proteins of Lactobacillus rhamnosus strains E/N, Oxy, and Pen, was performed. The obtained protein patterns allowed differentiation of the examined strains, which was not accomplished by the commonly used RAPD genotypic method. The differentiation by the SDS-PAGE method proved to be a useful tool for strain-specific identification, which was further confirmed by 2DE analysis. Therefore, it can be used as an alternative or complementary method for both conventional and genotypic identification procedures, especially when closely related lactobacilli isolates are identified.

• Preventive Nutrition and Food Science
• /
• v.7 no.1
• /
• pp.48-51
• /
• 2002

In order to study the reaction behavior of bovine holo- and apo-$\alpha$-lactalbumin ($\alpha$-La) during heat treatment at 65~10$0^{\circ}C$, the samples were analysed by first (ID)-and second-dimensional (2D) native-polyacrylamide gel electrophoresis (Native-PAGE) and sodium dodecylsulfate (SDS)-PAGE. When bolo-$\alpha$-La or apo- $\alpha$ -La were heated, they formed non-native, monomers, dimers and trimers. The apo-$\alpha$-La was more heat-sensitive than holo-$\alpha$-La. The monomers seemed to have the same composition as the native $\alpha$-La, but many of the disulfide bonds could be non-native.

• The Korean Journal of Zoology
• /
• v.35 no.1
• /
• pp.17-22
• /
• 1992

The three yolk polypeptides have been isolated and partially characterized. Their molecular weights of YPI, YP2, and YP3 were 48, 000, 47, 000, and 46, 000, respectivelv, as judged by SDS-polyacrvlamide gel electrophoresis. They have different digestion products upon in situ peptide mapping by limited proteolysis. Two-dimensional gel electrophoresis showed that their isoelectric points were heterogeneous from 5.92 to 6.54. And thew showed three different antigen-antibody reactions when each polvpeptides is reacted with antisera made to a mixture of all of three. These data reported here indicate that the yolk proteins are consisted of distinctive polypeptides in Drosophlla sp. (robusta species group).

• Journal of Life Science
• /
• v.13 no.3
• /
• pp.248-251
• /
• 2003

The kinetics of dimerization of a newly synthesized thyroglobulin (Tg), the precursor protein in the manufacture of thyroid hormone, was investigated in the endoplasmic reticulum of thyrocytes FRTL-5 cell line. The folded monomeric Tg was first detectable in a conformationally unstable form, from the examination of lysates of pulse labeled cultured thyrocytes by denaturing and nondenaturing gel electrophoresis by 15 min after biosynthesis. The first dimeric Tg was formed by 30 min after; the monomer declined and the dimer progressively increased, and 40 min after remarkable dimeric Tg form was found. Finally, dimerization was complete at 60 min after.

• Food Science of Animal Resources
• /
• v.26 no.2
• /
• pp.263-268
• /
• 2006

Probiotics including Lactobacillus acidophilus, refer to a group of nonpathogenic organisms that protect the human host against gastrointestinal(GI) infections by pathogenic bacteria such as Shiga toxin-producing E. coli(STEC). In the study, the inhibitory effects of STEC ATCC 43894 adhesion by L. acidophilus A4 was investigated on the HT-29 epithelial cells. Specific proteins regulated by cell Iysates of L. acidophilus A4 on STEC ATCC 43894 were also characterized by proteomic analysis. Both cell mass and Iysate of L. acidophilus A4 have exhibited the profound inhibitory activity on the HT-29 cells(about 1.5 log scale reduction). Two-dimensional gel electrophoresis(2-DE) revealed seven proteins that were up-regulated by cell Iysates of L. acidophilus A4 and three proteins that were down-regulated. In addition, three protein spots were only detected in the presence of cell Iysates. These results suggest that inhibitory effects of STEC adhesion by L. acidophilus may be due to the regulation of specific protein of STEC.