• The Korean Journal of Nuclear Medicine
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• v.33 no.6
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• pp.461-465
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• 1999

Scintillation proximity assay (SPA) is a unique type of radioimmunoassay and makes it possible to use radioisotopes for monitoring binding reactions continuously without separation procedure. Microbeads containing a fluorophor are covalently linked to antibody or receptor. When a radiolabeled antigen or ligand is added it binds to the beads and the emitted short range electrons, excite the fluorophor in the beads. The light emitted can be measured in a scintillation counter. $^3H$ or $^{125}I$ has been used for SPA. The sensitivities achieved with SPA are comparable to the sensitivities of other procedures. SPA is applicable to immunology, receptor binding, monitoring interactions of biomolecules and study for the kinetics of interaction between receptors and ligands.

• BMB Reports
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• v.41 no.3
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• pp.177-183
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• 2008

Ubiquitin is an essential, highly-conserved small regulatory protein in eukaryotic cells. It covalently modifies a wide variety of targeted proteins in the forms of monomer and polymers, altering the conformation and binding properties of the proteins and thus regulating proteasomal delivery, protein activities and localization. Mass spectrometry has emerged as an indispensable tool for in-depth characterization of protein ubiquitination. Ubiquitinated proteins in cell lysates are usually enriched by affinity chromatography and subsequently analyzed by mass spectrometry for identification and quantification. Ubiquitin-conjugated amino acid residues can be determined by unique mass shift caused by the modification. Moreover, the complex structure of polyubiquitin chains on substrates can be dissected by bottom-up and middle-down mass spectrometric approaches, revealing potential novel functions of polyubiquitin linkages. Here I review the advances and caveats of these strategies, emphasizing caution in the validation of ubiquitinated proteins and in the interpretation of raw data.

• Journal of Environmental Health Sciences
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• v.30 no.5 s.81
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• pp.374-377
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• 2004

Dual actions are the most recently used delivery system in drug study. Dual-action agents are unique chemical entities comprised of two different type of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. Crosslinked sulfadiazine-sulfanilamide such as antibiotics is synthesized by synthetic handle with glutaraldehyde. As a result, New synthetic antibacterial agent exhibited the broad antibacterial activities against Gram(+) and Gram(-) of 4 strains and a long durability supposing that the stomach and blood.

• Journal of the Korean Magnetic Resonance Society
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• v.21 no.1
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• pp.20-25
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• 2017

The cytoplasmic domains A and B of the mannitol transporter enzyme $II^{Mtl}$ are covalently linked in Escherichia coli, but separately expressed in Thermoanaerobacter Tengcongensis. The phosphorylation of domain B ($TtIIB^{Mtl}$) substantially increases the binding affinity to the domain A ($TtIIA^{Mtl}$) in T. Tengcongensis. To understand the structural basis of the enhanced domain-domain interaction by protein phosphorylation, we obtained NMR backbone assignments of the phospho-$TtIIB^{Mtl}$ using a standard suite of triple resonance experiments. Our results will be useful to monitor chemical shift changes at the active site of phosphorylation and the binding interfaces.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1280-1285
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• 2004

Retron is a prokaryotic genetic element, producing a short single-stranded DNA covalently linked to RNA (msDNA-RNA) by a reverse transcriptase (RT). In retron EC83, msDNA is further processed at between the 4th and the $5^{th}$ nucleotides, leaving a 79 nucleotide-long single-stranded DNA as a final product. To investigate this site-specific cleavage in msDNA synthesis, we purified the RT protein of retron EC83. Initially, RT ORF was cloned under the tac promoter, but the expression was very poor largely because of poor translation. In order to facilitate translation, the nucleotide sequence for the first nine amino acids was randomized with synonymous codons. This change of downstream sequence of translational initiation codon greatly affected the efficiency of translation. We could isolate clones which greatly increased RT production, and their sequences were compared to those of the low producers. The overproduced protein was purified and was shown to have RT activity.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.510.1-510
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• 1986

Many enzymes require the participation of readily dissociable coenzymes as NAD for thir catalytic activities. The continuous utilization of the enzymes requires the retention and regeneration of the coenzymes. For this purpose, several kinds of macromolecular NAD derivatives have been prepared by covalently attaching NAD to watersoluble polymers. We have prepared poly (ethylene glycol)-bound NAD (PEG-NAD) by coupling N$\^$6/-(2-carboxyethyl)-NAD to one terminal of ${\gamma}$ $\omega$-diaminoly (ethylene glycol) (Mr 3000) with water-soluble carbodiimide. PED-NAD thus obtained has one NAD moiety located at a terminal of the linear, flexible and hydrophilic chain of poly (ethylene glycol). PED-NAD has good coenzyme activity for various dehydrogenases and is applicable in a continuous enzyme reactor. To use these macromolecular NAD derivatives in an enzyme reactor, it si necessary to understand the behavior of the system in which the reactions of dehydrogenases are coupled by the recycling of the NAD derivative. We investigated the kinetic properties of a continuous enzyme reactor containing lactate dehydrogenase, alcohol dehydrogenase and PEG-NAD. The steady-state behavior of the enzyme reactor is explained by a simple kinetic model.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1254-1259
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• 2014

We investigated whether ${\beta}$-galactosidase (${\beta}$-gal)-containing Escherichia coli cells could transfer a galactose to 2-phenoxyethanol, resulting in 2-phenoxyethanol galactoside (PE-Gal). PE-Gal was confirmed by liquid chromatography-mass spectrometry. In addition, we also confirmed that a galactose molecule was covalently bonded with PE during thin-layer chromatography analysis of the ${\beta}$-gal hydrolysate of PE-Gal. The yield for PE-Gal synthesis was about 37.5% (weight basis), which was about 7-8 times greater than that of a previous report. In addition, the concentration of ${\beta}$-gal (0.96 U/ml) used in this PE-Gal synthesis was about 20 times less than that in a previous report.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.195-195
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• 2006

A superior electrophilic substitution reaction medium that is non-toxic, relatively less corrosive, and non-volatile electrophilic substitution reaction to afford high molecular weight linear and hyperbranched polyetherketones (PEK' s) was developed. The system has very strong driving force to give extra ordinary high molecular weight linear and hyperbranched PEK' s. The reaction medium was further extended to prepare various types of copolymers and covalently grafted polymers onto carbon nanotube (CNT) or carbon nanofiber (CNF). By using characteristic hydrophilic nature of the reaction medium, hyperbranched PEK' s could be synthesized from commercially available $A_3\;+\;B_2$ monomers without network formation via selective solubility of the monomers.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.378-378
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• 2006

The preparation, characterization, and bio-testing of biocide incorporated silicone coatings for marine applications have been conducted. Derivatives of the biocide, Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol), were used to covalently attach the biocide moiety to a silicone backbone. The synthetic process allowed for control of the resulting coating's mechanical properties as well as antifouling/fouling release performance in laboratory and ocean site testing. The test results showed significantly reduce macro fouling with sustained fouling release characteristics for the coatings produced.

• Bulletin of the Korean Chemical Society
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• v.26 no.9
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• pp.1381-1384
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• 2005

Surface modification of tridimensional cubic mesoporous silica, SBA-16, was investigated with pendant arm functionalized cobalt diaminosarcophagine (diAMsar) cage complex which covalently grafted onto the silica surface through the silication with sylanol group. The spectroscopic results showed that the mesoporous structure was preserved under the $[Co(diAMsar)]^{3+}$ grafting reaction condition. Successful grafting prevented the cobalt diAMsar cage from leaching out from the SBA-16 support.