• Development and Reproduction
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• v.22 no.1
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• pp.65-72
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• 2018

Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.10
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• pp.1484-1489
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• 2006

This study was carried out to investigate the effect of high pressure and the addition of non-meat proteins on the physico-chemical and binding properties of restructured pork. Pressurizations were carried out at up to 200 MPa and non-meat proteins used as a binder were isolated soy protein (ISP), sodium caseinate (SC), whey protein concentrate (WPC) and egg white powder (EWP). The pH values of all treatments were affected by the level of pressure. $L^*$-value of all treatments increased significantly (p<0.05), while both $a^*$-value and $b^*$-value of all treatments showed a significant decrease (p<0.05) with increasing pressure level. Binders could contribute only additive effects on both pH and color of the treatments. It was found that high pressure improved the water binding capacities and binding strength of the treatments. Binders also improved the binding strength of restructured pork. However, SC and WPC had no effect on water binding properties under high pressure. These results indicate that the application of high pressure had more significant effect on restructuring meat than binders.

• Food Science and Preservation
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• v.19 no.3
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• pp.438-442
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• 2012

To prepare calcium-binding peptides as calcium supplement, barley proteins were hydrolyzed using Flavourzyme for 18 h and the hydrolysates were ultra-filtered under 3 kDa as a molecular weight. The resultant filtered peptides were fractionated using ion exchange and normal-phase high performance liquid chromatography. Then each fraction that was obtained was determined for its calcium-binding activity to isolate the calcium-binding peptides. As a result, the highest calcium-binding peptide fraction was obtained, and the results suggest that barley protein hydrolysates can be used as a calcium supplement.

• Journal of the Korean Magnetic Resonance Society
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• v.25 no.2
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• pp.17-23
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• 2021

High mobility group protein B1 (HMGB1) is a highly conserved, non-histone, chromatin associated nuclear protein encoded by HMGB1 gene. HMGB1 proteins may be general co-factors in Hox-mediated transcriptional activation that facilitate the access of Hox proteins to specific DNA targets. It is unclear that the exact binding interface of Hoxc9DBD and HMGB1. To identify the interface and binding affinity of Hoxc9DBD and HMGB1 A-box, the paramagnetic probe, MTSL was used in NMR titration experiment. It is attached to the N-terminal end of HMGB1 A-box by reaction with thiol groups. The backbone assignment of HMGB1 A-box was achieved with 3D NMR techinques. The ¹⁵N-labeled HMGB1 A-box was titrated with MTSL-labeled Hoxc9DBD respectively. Based on the chemical shift changes we can identify the interacting residues and further map out the binding sites on the protein structure. The NMR titration result showed that the binding interface of HMGB1 A-box is around loop-1 between helix-1 and helix-2. In addition, the additional contacts were found in N- and C-terminus. The N-terminal arm region of Hoxc9DBD is the major binding region and the loop between helix1 and helix2 is the minor binding region.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.81.2-82
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• 2003

Plants have evolved along with pathogens, and they have developed sophisticated defense systems against specific microorganisms to survive. G-protons are considered one of the upstream signaling components working as a key for the defense signal transduction pathway. For activation and inactivation of G-protein, GTP-biding proteins are involved. GTP -binding proteins are found in all organisms. Small GTP-binding proteins, having masses of 21 to 30kD, belong to a superfamily, often named the Ras supefamily because the founding members are encoded by human Ras genes initially discovered as cellular homologs of the viral ras oncogene. Members of this supefamily share several common structural features, including several guanine nucleotide binding domains and an effector binding domain. However, exhibiting a remarkable diversity in both structure and function. They are important molecular switches that cycle between the GDP-bound inactive form into the GTP-bound active form through GDP/GTP replacement. In addition, most GTP-binding proteins cycle between membrane-bound and cytosolic forms. such as the RAC family are cytosolic signal transduction proteins that often are involved in processing of extracellular stimuli. Plant RAC proteins are implicated in regulation of plant cell architecture secondary wall formation, meristem signaling, and defense against pathogens. But their molecular mechanisms and functions are not well known. We isolated a RacB homolog from rice to study its role of defense against pathogens. We introduced the constitutively active and the dominant negative forms of the GTP-hinging protein OsRacB into the wild type rice. The dominant negative foms are using two forms (full-sequence and specific RNA interference with RacB). Employing southern, and protein analysis, we examine to different things between the wild type and the transformed plant. And analyzing biolistic bombardment of onion epidermal cell with GFP-RacB fusion protein revealed association with the nucle.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1088-1097
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• 2021

Grouper nervous necrosis virus (GNNV) infection causes mass grouper mortality, leading to substantial economic loss in Taiwan. Traditional methods of controlling GNNV infections involve the challenge of controlling disinfectant doses; low doses are ineffective, whereas high doses may cause environmental damage. Identifying potential methods to safely control GNNV infection to prevent viral outbreaks is essential. We engineered a virus-binding bacterium expressing a myxovirus resistance (Mx) protein on its surface for GNNV removal from phosphate-buffered saline (PBS), thus increasing the survival of grouper fin (GF-1) cells. We fused the grouper Mx protein (which recognizes and binds to the coat protein of GNNV) to the C-terminus of outer membrane lipoprotein A (lpp-Mx) and to the N-terminus of a bacterial autotransporter adhesin (Mx-AIDA); these constructs were expressed on the surfaces of Escherichia coli BL21 (BL21/lpp-Mx and BL21/Mx-AIDA). We examined bacterial surface expression capacity and GNNV binding activity through enzyme-linked immunosorbent assay; we also evaluated the GNNV removal efficacy of the bacteria and viral cytotoxicity after bacterial adsorption treatment. Although both constructs were successfully expressed, only BL21/lpp-Mx exhibited GNNV binding activity; BL21/lpp-Mx cells removed GNNV and protected GF-1 cells from GNNV infection more efficiently. Moreover, salinity affected the GNNV removal efficacy of BL21/lpp-Mx. Thus, our GNNV-binding bacterium is an efficient microparticle for removing GNNV from 10‰ brackish water and for preventing GNNV infection in groupers.

• Journal of Nutrition and Health
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• v.32 no.6
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• pp.628-636
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• 1999

There is a marked increase in geriatric disease, especially liver disease, due to the continuous increase in alcohol and fat consumption. Since the fatty liver, induced by alcohol or fat, is basically from abnormalities in the lipid metabolism, it is possible that fatty acid binding protein(FABP) which is related to the fatty acid metabolism may also be abnormal in these livers. FABP is a small molecular weight protein family present in cytosol in high concentration. It has been proposed as a fatty acid transfer protein and as a binding protein responsible for controlling intracellular free fatty acid concentration. In this research, we have examined the relationship between liver FABP and fatty liver induced by alcohol or high cholesterol diet. Rats were fed one of either semipurified liquid diets; control diet containing 65% carbohydrate, 20% protein, and 15% fat or high cholesterol diet containing 1%(w/w) cholesterol or alcohol diet containing 37% of alcohol instead of carbohydrate. After 5 weeks of feeding period, all rats received commercial chow diet for 5 weeks to examine recovery effect. Liver and blood samples were collected at 0, 1, 3, 5 and 10 weeks to analyze lipid compositions. FABP was purified from liver cytosol and injected to rabbit to obtain antiserum. Liver FABP amount was determined by SDS-PAGE and western blotting methods. Fatty acid binding capacity was determined by binding of 14Cpalmitate with the delipidated liver cytosol. Consumption of alcohol increased serum cholesterol, triglyceride concentration and decreased HDL-cholesterol concentration after 5 weeks. Serum apolipoprotein B concentration increased after 3 weeks and LDL-cholesterol and apolipoprotein A concentration changed after 1 week. Liver cholesterol and triglyceride concentration increased after 3 weeks. Consumption of high cholesterol diet changed liver and serum lipid composition after 3 weeks. Swiching to normal diet for 5 weeks did not normalize most of lipid composition in serum and liver except serum and liver except serum cholesterol, triglyceride and liver cholesterol. Liver cytosol FABP content and the fatty acid binding capacity decreased dramatically after 1 week with alcohol consumption. This results indicate that FABP content changes before the changes before the changes of blood or liver lipid composition, suggesting changes of FABP may cause development of the fatty liver induced by alcohol and can be used as an index of detecting a early development of fatty liver.

• Archives of Pharmacal Research
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• v.10 no.1
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• pp.29-35
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• 1987

The effect of drug addition on the bovine serum albumin (BSA)-edible dye complex was studied by spectrophotometric method. The edible dyes tested were amranth, erythrosine, tatrazine and sunset yellow. The moles of bound dye per protein mole and free energies for edible dyes bounded were determined at pH 7.4. The values of free energy change by the addition of drughs to BSA-edible dye were ranged fro -6, 260 to 08030 cal/mole. In the wide range of edible dye concentration (0.3-$7{\times}10^{-5}$$^{-5}$ M), acetylsalicylic acid (ASA) showed pattern of displacement different from that of dye. It was assumed that ASA has different binding mechanisms from edible dye.

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.238-244
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• 1993

We identified juvenile hormone binding protein (JHBP) from last instar larval hemollvnph of Hvphantria cunea using gel filtration and non-SDS PAGE. Two kinds of JHBP in hemollnnph were found at two peaks by gel filtration (Sephadex G-100) and also at Rm values of 0.13 and 0.57 by non-SDS PAGE. JHBP was partially purified using anion exchange chromatosraphv, preparative gel filteration, and preparative PAGE. Dextrin coated charcoal (DCC) binding assay was employed to monitor the location of JHBP in chromatographic profile during the purification process. Purity of JHBP was checked by silver staining of 1091 SDS-Polyacrvlamide.

• Journal of Photoscience
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• v.3 no.2
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• pp.85-90
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• 1996

The binding ability of leaf nuclear extracts to the lighbresponsive elements (LREs) of cab promoters of Arabidopsis thaliana has been investigated. The cab promoters were fragmented with restr ction endonucleases into LRE that were identified by Mitra et al. [Plant Mol. Biol. 12, 169179 ( 1989)] and other small fragments. After end labeling with Klenow fragment, the fragments were assayed for binding with the leaf nuclear proteins that were prepared by solubilizing the purified nuclei with 0.5 M ammonium sulfate. The binding ability was assayed by mobility shift assay. To perform successful mobility shift assay, several factors affecting the interaction of protein with DNA were optimized before performing the assay. The LREs had several retardation bands. However, the other promoter fragments from the transcription start site to the far upstream region of the promoters had also retardation bands. No particular relationships could be found between the retardation band distributions and the loci of LRE. It is likely that the light-regulation of cab gene expression may be controlled by the multiple interactions of the regulatory protein factors with DNA motifs.