• BMB Reports
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• 제30권4호
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• pp.296-298
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• 1997

A bacterial glutathione S-transferase from Pseudomonas sp. DJ77 has been crystallized. The crystals diffract to at least $2.3\;\AA$ resolution, and belong to the orthorhombic space group $P2_{1}2_{1}2_{1}$, with cell parameters $a=97.4\;\AA,\;b=100.3\;\AA$, and $c=46.0\;\AA$. There is one dimer molecule of pGST per crystallographic asymmetric unit. with the crystal volume per protein mass of $2.34\;\AA^3/dalton$ and a solvent content of about 47% (v/v).

• 대한화학회지
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• 제67권5호
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• pp.348-352
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• 2023

p97, a universally conserved AAA+ ATPase, holds a central position in the ubiquitin-proteasome system, orchestrating myriad cellular activities with significant therapeutic implications. This protein primarily interacts with a diverse set of adaptor proteins through its N-terminal domain (NTD), which is structurally located at the periphery of the D1 hexamer ring. While there have been numerous structural elucidations of p97 complexed with adaptor proteins, the stoichiometry has remained elusive. In this work, we present the crystal structure of the p97-N/D1 hexamer bound to the FAF1-UBX domain at a resolution of 3.1 Å. Our findings reveal a 6:6 stoichiometry between the p97 hexamer and FAF1-UBX domain, deepening our understanding from preceding structural studies related to p97-NTD and UBX domain-containing proteins. These insights lay the groundwork for potential therapeutic interventions addressing cancer and neurodegenerative diseases.

• Bulletin of the Korean Chemical Society
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• 제31권5호
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• pp.1223-1227
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• 2010

In this study, the orientational behavior of thermotropic liquid crystals (LC) supported on a film of protein receptors was examined. Avidin was roller printed and covalently immobilized onto the surface of gold using NHS/EDC chemistry. The orientation of nematic 4-cyano-4'-pentylbiphenyl (5CB) was found to be parallel to the plane of the printed avidin surface before incubation with a solution of biotin. However, protein-receptor complexation induced a random orientation of 5CB, where protein-receptor complexes disturbed the nanoscale topography of the printed protein surface. Atomic force microscopy and ellipsometry was used to confirm printing and the specific interaction of proteins. These results demonstrate that the combination of LC and roller printing can be used to detect specific interactions between biomolecules by manipulating the orientational behavior of LC to the printed protein surfaces.

• Applied Science and Convergence Technology
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• 제24권1호
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• pp.1-8
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• 2015

Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.639-642
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• 2003

본 연구는 후각 수용 단백질인 ODR10를 GST와 Histidine tag를 각각 N 말단과 C 말단에 삽입한 후 두 가지의 발현 벡터에 넣어 대장균에서 발현시켰다. 부분 정제된 단백질을 QCM의 수정진동자에 코팅한 후 여러 종류의 냄새 분자와의 상호 작용을 관찰하였다. 발현양은 적었지만 QCM실험 결과 발현된 단백질이 diacetyl과 반응한다는 것을 알 수 있었다. ODR10 단백질과 diacetyl의 결합 정도는 다른 냄새 분자와 비교했을 때 $5{\sim}10$배 정도 차이가 났으며 이를 통해 후각 수용 단백질을 발현시킨 대장균 세포들을 후각센서를 개발하는데 사용할 수 있다는 것을 알 수 있었다. 또한 현재까지는 1000가지 이상 존재한다고 알려진 후각 수용 단백질들이 어떤 냄새 분자와 특이적인 결합성을 가지는지 조사하기 위해서는 복잡하고 시간이 오래 걸리는 실험을 해야 했었지만, 대장균에서 발현시키는 시스템을 통해 경제적이고 효율적으로 조사를 할 수 있게 되었다.

• 식물조직배양학회지
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• 제27권4호
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• pp.339-343
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• 2000

When plants are exposed to subfreezing temperatures ice crystals are forming within extracelluar space in leaves. The growth of ice crystal is closely related to the degree of freezing injury. It was shown that an antifreeze protein binds to an ice nucleator through hydrogen bonds to prevent growth of ice crystal and also reduces freezing damage. The antifreeze proteins in plants are similar to PR proteins but only the PR proteins induced upon cold acclimation were shown to have dual functions in antifreezing as well as antifungal activities. Three of the genes encoded for CLP, GLP, and TLP were isolated from barley and Kentucky bluegrass based on amino acid sequence revealed after purification and low temperature-inducibility as shown in analysis of the protein. The deduced amino acid of the genes cloned showed a signal for secretion into extracellular space where the antifreezing activity sup-posed to work. The western analysis using the antisera raised against the antifreeze proteins showed a positive correlation between the amount of the protein and the level of freeze tolerance among different cultivars of barely. Besides it was revealed that TLP is responsible for a freeze tolerance induced by a treatment of trinexapac ethyl in Kentucky bluegrass. Analysis of an overwintering wild rice, Oryza rufipogon also showed that an acquisition of freeze tolerance relied on accumulation of the protein similar to CLP. The more direct evidence for the role of CLP in freeze tolerance was made with the analysis of the transgenic tobacco showing extracellular accumulation of CLP and enhanced freeze tolerance measured by amount of ion leakage and rate of photosynthetic electron transport upon freezing. These antifreeze proteins genes will be good candidates for transformation into crops such as lettuce and strawberry to develop into the new crops capable of freeze-storage and such as rose and grape to enhance a freeze tolerance for a safe survival during winter.

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.389-392
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• 1988

Escheriachia coli pSL 2-1 clone은 Bacillus sphaericus 1593의 모기살충 유전자를 클로닝한 재조합 DNA이다. 이 클론이 생산하는 살충독소 단백질의 분자량을 SDS-polyacrylamide gel을 이용하여 측정했다. B. sphaericus 1593균이 생산하는 독소결정체를 분리하여 전기영동을 한 결과는 6개의 단백질밴드(43, 58, 64, 100, 113, 130Kd)가 형성되었으나, 독소결정체를 알칼리 pH로 용해하여 전기영동을 하면 2개의 단백질 밴드(43과 64Kd)만이 나타났다. 그러나 대장균 pSL2-1균이 생산하는 독소단백질을 Sephadex G-200으로 정제하여 모기유충에 살충력이 있는 단백질을 전기영동한 결과는 42Kd만이 나타났다. LC50은 2 $\mu\textrm{g}$/$m\ell$이었다. B. sphaericus와 pSL2-1 clone 생산하는 살충단백질은 42Kd 단백질인 것으로 생각된다.

• 한국응용곤충학회지
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• 제33권3호
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• pp.178-183
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• 1994

Bacillus thuringiensis kurstaki와 aizawai의 내독소단백질을 알카리용액 또는 트립신, 곤충소화액 드응ㄹ 처리하여 전기영동한 후 단백질팬턴을 비교하였다. 두 균주의 주요 결정단백질은 130kd와 64kd의 단백질이었으며, 소화액이나 효소로 처리한 경우 공통적으로 62kd의 활성독소가 생성되었다. 그러나, aizawai는 kurstaki에 비해 현저히 적은 양의 62kd 단백질을 생성하였다. 흰불나방의 유충이 Bacillus thuringiensis 독소를 섭식하였을 때 지방체를 비롯한 몇 가지 조직에서 45kd의 스트레스 단백질이 유발되었는데 이 단백질은 열충격이나 저온 충격시에도 마찬가지로 생성되었다.

• 한국잠사곤충학회지
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• 제35권1호
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• pp.36-42
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• 1993

Bacillus thruingiensis subsp. kurstaki HD-1의 내독소단백질 유전자인 CryIA(a)의 N-말단 부분과 B.tk HD-73의 내독소단백질 유전자인 CryIA(c)의 C-말단 부분의 융합에 따른 독성발현 여부에 관하여 조사하였다. 융합생산물인 pSK3, pSK4 및 pSK5의 플라스미드는 각각 4.5kb, 4.8kb 그리고 5.5kb로 구성되어 있다. 형질전환체의 내독소단백질에 대한 Western blotting은 SK4 및 SK5가 77-kDa 그리고 105-kDa에서 B.t k HD-1 항체에 대하여 반응을 나타내었다. 독성검정의 공시충인 배추좀나방 및 담배나방을 사용한 결과 pSK5를 포함하는 형질전환체만이 각각 96% 및 97%의 높은 치사율을 나타냈다.

• Biodesign
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• 제7권2호
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• pp.35-37
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• 2019

Xanthomonas oryzae pv. oryzae (Xoo) is a plant pathogen, which causes a bacterial blight of rice. The bacterial blight is one of the most devastating diseases of rice in most of the rice growing countries and there is no effective pesticide against bacterial blight. The β-ketoacyl-acyl carrier protein synthase III (FabH) plays a key role in fatty acid synthesis (FAS) and is a promising drug target for the development of antibacterial agents. Xoo0878 gene, a fabH gene, from Xoo was cloned and its gene product Xoo0878 was expressed, purified and crystallized. Xoo0878 crystal diffracted to 2.1Å resolution and belonged to the triclinic space group P1, with unit-cell parameters a = 57.3Å, b = 64.7Å, c = 104.2Å and α = 81.6°, β = 84.7°, γ = 74.4°. There are four monomers in the asymmetric unit, with a corresponding crystal volume per protein weight of 2.65 ų Da^-1 and a solvent content of 53.6%. Xoo0878 structure will be useful to develop new antibacterial agents against Xoo.