• BMB Reports
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• v.46 no.12
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• pp.606-610
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• 2013

Glucagon like peptide-1 (GLP-1) regulates glucose mediated-insulin secretion, nutrient accumulation, and ${\beta}$-cell growth. Despite the potential therapeutic usage for type 2 diabetes (T2D), GLP-1 has a short half-life in vivo ($t_{1/2}$ <2 min). In an attempt to prolong half-life, GLP-1 fusion proteins were genetically engineered: GLP-1 human serum albumin fusion (GLP-1/HSA), AGLP-1/HSA which has an additional alanine at the N-terminus of GLP-1, and AGLP-1-L/HSA, in which a peptide linker is inserted between AGLP-1 and HSA. Recombinant fusion proteins secreted from the Chinese Hamster Ovary-K1 (CHO-K1) cell line were purified with high purity (>96%). AGLP-1 fusion protein was resistant against the dipeptidyl peptidase-IV (DPP-IV). The fusion proteins activated cAMP-mediated signaling in rat insulinoma INS-1 cells. Furthermore, a C57BL/6N mice pharmacodynamics study exhibited that AGLP-1-L/HSA effectively reduced blood glucose level compared to AGLP-1/HSA.

• Journal of fish pathology
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• v.22 no.2
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• pp.173-177
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• 2009

Lymphocystis disease virus (LCDV) was detected from olive flounder Paralichthys olivaceus, painted glass fish Chanda baculis, gourami Trichogaster leeri and rockfish Sebastes schlegeli, and proteins of the viruses were compared. The major capsid protein (MCP) gene-specific primer sets successfully amplified approximately 1300 bp nucleotides from the olive flounder and 600 bp nucleotides from painted glass fish, gourami and rockfish isolates, respectively. In western blotting analysis using anti-LCDV mouse polyclonal serum, major antigenic proteins had 21, 26, 45, 50, 80, 110 and 120 kDa in olive flounder, 26, 47 and 80 kDa in painted glass fish, 26, 46, 80 and 92 kDa in gourami, 26, 44, 49, 80 and 105 in rockfish, respectively. All the marine and freshwater isolates showed only common antigens of approximately 26 kDa and 80 kDa. These results suggest that antigenic protein profiles of LCDVs may vary depending upon fish species.

• Mass Spectrometry Letters
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• v.12 no.3
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• pp.76-80
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• 2021

Scrub typhus is an acute febrile disease caused by the pathogenic bacterium Orientia tsutsugamushi, belonging to the Rickettsiaceae family. The shotgun proteomic analysis was performed using the sera of scrub typhus patients to identify the proteins having their origin in O. tsutsugamushi. Three different databases approaches were used for the identification of the proteomes. We identified the RsmD, an RNA methyltransferase as the commonly detected protein from all three approaches. This protein was not detected in the sera of healthy negative controls. We believe that this protein is a potential biomarker of Orientia tsutsugamushi present in the sera of scrub typhus patients.

• Journal of fish pathology
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• v.12 no.2
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• pp.89-99
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• 1999

New sixteen heat shock proteins (Hsps) and ten ethanol shock proteins were appeared on the analysis with SDS-PAGE when cultivation temperature for the Vibrio vulnifrcus ATCC 27562 strain was shifted-up to $42^{\circ}C$ from $30^{\circ}C$ for 20 mins and treated with of 6% ethanol for 10 mins, respectively. Even the induction of thermotolerance in V. vulnificus was coincided with the induction of Hsps if the pre-shock was adjusted to thermal temperature. Outer membrane proteins (OMPs) that were purified from the membrane of cells after heat shock showed more immunodominant pattern to the immunized rabbit anti-V. vulnificus O serum in enzyme-linked immunosorbent assay (ELISA). On the western immunoblot analysis it was confirmed that both 62 kDa IMP and 69 kDa OMP in the Hsps and 48 kDa IMP a major OMP in the ethanol shock proteins were reacted with rabbit anti-V. vulnificus O sera. Agglutination titer of the heat shocked V. vulnificus with rabbit anti-V. vulnificus O serum was higher than that of the untreated bacteria.

• Applied Biological Chemistry
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• v.31 no.4
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• pp.361-370
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• 1988

To investigate the interaction between whey and soybean protein, thermal changes of component proteins were analyzed by column chromatography and gel electrophoresis. In the Sephadex G-200 chromatography of the mixture treated at above $80^{\circ}C$, the amount of low molecular weight proteins and high molecular aggregates were increased. This implicated that dissociation of 1ls globulin into subunits and the formation of soluble aggregates between these subunits and whey proteins that contain thiol and disulfide groups. These interaction between soy proteins and ${\beta}-lactoglobulin$, ${\alpha}-lactalbumin$, and proteose-peptone 3 were confirmed by gel electrophoresis. Bovine serum albumin, Immunoglobulin-G(H), Lactoferrin, 1ls-subunits(basic and acidic), and subunit of 7s globulin were also considered to interact each other depending on the condition of the salt solutions.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.6
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• pp.893-901
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• 2017

Objective: Hypocalcemia is an important metabolic disease of dairy cows during the transition period, although the effect of hypocalcemia on biological function in dairy cows remains unknown. Methods: In this study, proteomic, mass spectrum, bioinformatics and western blotting were employed to identify differentially expressed proteins related to serum Ca concentration. Serum samples from dairy cows were collected at three time points: 3rd days before calving (day -3), the day of calving (day 0), and 3rd days after calving (day +3). According to the Ca concentration on day 0, a total of 27 dairy cows were assigned to one of three groups (clinical, subclinical, and healthy). Samples collected on day -3 were used for discovery of differentially expressed proteins, which were separated and identified via proteomic analysis and mass spectrometry. Bioinformatics analysis was performed to determine the function of the identified proteins (gene ontology and pathway analysis). The differentially expressed proteins were verified by western blot analysis. Results: There were 57 differential spots separated and eight different proteins were identified. Vitamin D-binding protein precursor (group-specific component, GC), alpha-2-macroglobulin (A2M) protein, and apolipoprotein A-IV were related to hypocalcemia by bioinformatics analysis. Due to its specific expression (up-regulated in clinical hypocalcemia and down-regulated in subclinical hypocalcemia), A2M was selected for validation. The results were consistent with those of proteomic analysis. Conclusion: A2M was as an early detection index for distinguishing clinical and subclinical hypocalcemia. The possible pathogenesis of clinical hypocalcemia caused by GC and apolipoprotein A-IV was speculated. The down-regulated expression of GC was a probable cause of the decrease in calcium concentration.

• Proceedings of the KIEE Conference
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• 2002.11a
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• pp.60-63
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• 2002

This paper presents bead-based microbiocihps to detect and separate target proteins. Micro beads coated with capture proteins were introduced into a microchamber, and target proteins flowing across the chamber were bound and concentrated. The chip was connected with an external fluid system. Bead surfaces were double-coated with photo-cleavable linkers and capture proteins. The proteins bound on the beads were photo-separated under UV irradiation, and excited to be measured in fluorescence. $38{\sim}50{\mu}m$ sized polystyrene beads were used. SOGs(silicon-on-glass) were used to fabricate the microchip having glasses bonded on both sides. 100 ${\mu}m$ thick silicon channel was formed through silicon deep RIE process. The upper glass cover had holed through to have inlets and outlets fabricated by powder-blastings. In this study, biotin and streptavidin were used as capture proteins and detection proteins, respectively. The protein mixtures of streptavidin, HSA(human serum albumin) and ovalbumin were applied for selective detection test.

• Journal of Chest Surgery
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• v.25 no.11
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• pp.1209-1217
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• 1992

Triiodothyronine[T3] is an important regulator of the tissue metabolism, and may have potential use as an inotropic agent. The change of serum T3 level was studied in the pediatric age patients after cardiopulmonary by pass. Thyroid function was tested pre-operatively in 33 patients and total triiodothyronine[TT3] levels were serially measured during and after cardiopulmonary bypass[CPB]. After correction of dilutional effect, effects of various factors on the TT3 levels were analyzed. Abrupt fall of TT3 level was demonstrated at 15 minutes after CPB[80.1$\pm$5.9ng/dL] from the initial level of 133.6$\pm$5.3ng/dL, with some recovery at 6 hours[115.4$\pm$6.7ng/dL]. After then, gradual decrease occured reaching to the level of 77.2$\pm$4.2ng /dL at 24 hours. These falls of the TT3 after CPB were statistically significant. [p<0.01 ANOVA] Statistically significant correlation were found between the degree of hemodilution and TT3 concentration at 15 and 30 minutes after CPB[p<0.05]. But, other factors were analyzed to have no effect on TT3 levels. As the degree of the hemodilution increases, TT3 decreased less. This observation probably supports the fact that decrease of TT3 during CPB may be a result of sequestration of T3 into peripheral tissue. Although it was not statistically significant[p=0.08], the fall of TT3 was greater in the group to which plasmanate was added, than those not added. This finding seemed to be due to the increase of albumin and other thyroid-hormone-binding-proteins in the serum. Increase of these binding proteins might potentiate the sequestration of T3 into the liver and the kidney from serum, and as a consequence, decrese the serum TT3 level further.

• BMB Reports
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• v.33 no.2
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• pp.133-137
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• 2000

To elucidate the effect of oxygen radicals on the molecular properties of proteins, the secondary and tertiary structure and molecular weight size of BSA and ${\beta}$-lactoglobulin were examined after irradiation of proteins at various doses. Gamma-irradiation of protein solutions caused the disruption of the ordered structure of protein molecules as well as degradation, cross-linking, and aggregation of the polypeptide chains. As a model system, BSA and ${\beta}$-lactoglobulin were used as a typical ${\alpha}$-helical and a ${\beta}$-sheet structure protein, respectively. A circular dichroism study showed that the increase of radiation decreased the ordered structure of proteins with a concurrent increase of aperiodic structure content. Fluorescence spectroscopy indicated that irradiation quenched the emission intensity excited at 280 nm. SDS-PAGE and a gel permeation chromatography study indicated that radiation caused initial fragmentation of proteins resulting in a subsequent aggregation due to cross-linking of protein molecules.

• Macromolecular Research
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• v.17 no.4
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• pp.232-239
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• 2009

This paper describes the simple nanopatterning of proteins on polyelectrolyte surfaces using microcontact printing with a nanopatternable, hydrophilic composite nanomold. The composite nanomold was easily fabricated by blending two UV-curable materials composed of Norland Optical Adhesives(NOA) 63 and poly(ethylene glycol) dimethacrylate(PEG-DMA). NOA 63 provided stable nanostructure formation and PEG-DMA induced high wettability of proteins in the nanomold. Using the composite mold and functionalized surface with polyelectrolytes, the fluorescent, isothiocyanate-tagged, bovine serum albumin(FITC-BSA) was successfully patterned with 8 nm height and 500 nm width. To confirm the feasibility of the protein assay on a nanoscale, a glycoprotein-lectin assay was successfully demonstrated as a model system. As expected, the lectins correctly recognized the nano-patterned glycoproteins such as chicken ovalbumin. The simple preparation of composite nanomold and functionalized surface with a universal platform can be applied to various biomolecules such as DNA, proteins, carbohydrates, and other biomolecules on a nanoscale.