• Mass Spectrometry Letters
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• 제2권2호
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• pp.53-56
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• 2011

[ $C_8$ ]functionalized magnetic nanoparticles were synthesized by coating magnetic $Fe_3O_4$ nanoparticles with silicaamine groups using 3-aminopropyltriethoxysilane and by subsequently modifying the amine groups with chloro(dimethyl)octylsilane to produce octyl groups on the surface of the MNPs. The $C_8$-functionalized MNPs were used to enrich peptides from tryptic protein digests of myoglobin and ${\alpha}$-casein. The enriched peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS was also used to investigate desalting of the $C_8$-functionalized MNPs. Sample solutions were prepared in 1.0 M NaCl, and the successful removal of salt was observed. Enrichment with $C_8$-functionalized MNPs was very effective for separating and concentrating tryptic peptides.

• 한국식품위생안전성학회지
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• 제32권4호
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• pp.284-289
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• 2017

본 연구는 국내 미지정 피막제인 산화형 폴리에틸렌 왁스를 분석하기 위해 다양한 분석방법을 비교, 검토하였으며, 확립된 최적 분석법을 이용하여 시중 유통되는 과일 및 견과류에서 산화형 폴리에틸렌 왁스의 사용여부를 모니터링 하였다. 최적 분석법을 확립하기 위하여 산가, HT-GPC, MALDI-TOF/MS, GC-FID, FT-IR 등을 비교하였다. 산가측정 결과 산화형 폴리에틸렌 왁스를 첨가한 시료와 첨가하지 않은 시료 모두 산가가 1.12 mg KOH/g로 산가의 변화가 나타나지 않아 산가를 이용한 식품 중 산화형 폴리에틸렌 왁스의 분석은 어려울 것으로 판단되었으며, HT-GPC분석은 표준물질의 분산도 값이 약 3.4로 확인되었으며, GPC를 이용한 분석은 실험실간 낮은 재현성과 컬럼 등 분석조건에 따른 결과 값이 달라질 수 있기 때문에 분자량의 분산도만으로 식품 중 산화형 폴리에틸렌 왁스의 분석은 어려울 것으로 판단되었다. MALDI-TOF/MS 분석은 700~2,000 m/z에 peak가 관찰되었지만, 산화형 폴리에틸렌 왁스 성적서에 적혀있는 분자량 범위인 2,700~2,800 m/z와 큰 차이를 보여, MALDI-TOF/MS를 이용한 분석 역시 어려울 것으로 판단되었다. GC-FID를 이용한 지방산 분석결과 산화형 폴리에틸렌 왁스 표준물질에서 과일 및 견과류에서 흔히 볼 수 없는 지방산인 C11:0, C15:1 지방산이 검출되었지만, 제외국 허용기준으로 첨가하여 분석하였을 때 관찰되지 않고, 고농도를 첨가할 경우에만 피크가 관찰되어 GC-FID를 이용한 분석은 어려울 것으로 판단되었다. FT-IR 분석결과 코팅된 견과류 시료와 코팅되지 않은 견과류 시료를 비교하였을 때 코팅된 시료에서 산화형 폴리에틸렌 왁스 표준물질의 spectrum과 일치하는 spectrum을 나타내어, 식품 중 산화형 폴리에틸렌 왁스의 분석은 FT-IR로 분석이 가능함을 확인하였다. 확립된 FT-IR 분석법을 이용하여 국내외 대형마트에서 구입한 111건의 과일 및 견과류를 분석한 결과 산화형 폴리에틸렌 왁스를 사용한 시료가 없음을 확인하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.311-314
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• 2003

Under hyperosmotic stress, rCHO cells display decreased specific growth rate $({\mu})$ and increased specific antibody productivity $(q_{Ab})$. The effects of hyperosmotic stress on batch culture cellular dynamics are not well understood. To this end, we conducted a proteome profile of rCHO cells, using 2D-gel, MALDI-TOF-MS and MS/MS. As a result, the proteome profile of rCHO cells could be established using 41 identified proteins. Based on this proteome profile of rCHO cells, we have found at least 8 differently expressed spots at hyperosmotic osmolality (450 mOsm/kg). Among these spots, two metabolic enzymes were found to be up-regulated (pyruvate kinase and GAPDH), while down-regulated protein was identified as tubulin. It shows that hyperosmotic stress can alter metabolic state, by up-regulated activities of two glycolysis enzymes, which could lead to activate the generation of metabolic energy. Tubulin expression was down-regulated, suggesting a reduction of cell division. Finally, the increased conversion energy could leads to improve overall productivity.

• 대한임상검사과학회지
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• 제50권3호
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• pp.275-283
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• 2018

Helicobacter pylori는 만성 위염, 위궤양 또는 위 선암을 비롯한 다양한 위장병의 원인균으로 알려져 있으며, 이 세균이 분비하는 cytotoxin-associated protein A (CagA), vacuolating cytotoxic protein A (VacA)와 같은 병원성 인자가 그 원인으로 알려져 있다. 본 연구에서는 zerumbone이 CagA, VacA를 비롯한 다양한 H. pylori의 병원성 인자들의 단백 발현양 변화에 정성적, 정량적으로 어떠한 영향을 미치는지 분석하였다. H. pylori 60190 (VacA 양성 / CagA 양성; Eastern type) 표준균주를 대상으로 하여 약 200개의 유의미한 단백을 선별한 후, 그 중에서 임상적으로 의의가 큰 13개 단백 분자에 대한 프로테옴 분석을 수행하였다. 이차원 전기 영동법(2 dimensional electrophoresis, 2-DE) 수행 후, 단백질 스팟의 정량적 측정에는 ImageMasterTM 2-DE Platinum 소프트웨어를 사용하였고, 단백 동정에는 matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF-MS)와 liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS)를 사용하였다. 동정이 완료된 전체 단백 중에서 유의미한 변화를 보인 단백을 집중적으로 분석한 뒤에 필요에 따라 reverse transcription -polymerase chanin reaction을 수행하여 결과를 추가 검증하였다. 본 연구에서는 zerumbone이 보유한 새로운 약리학적 활성 기전을 스크리닝함으로써 향후 zerumbone이 H. pylori 감염증 치료제로서 어떠한 의미를 지니는지 규명하고자 하였다.

• Parasites, Hosts and Diseases
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• 제54권1호
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• pp.71-74
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• 2016

Trichomonas vaginalis is a flagellate protozoan parasite and commonly infected the lower genital tract in women and men. Iron is a known nutrient for growth of various pathogens, and also reported to be involved in establishment of trichomoniasis. However, the exact mechanism was not clarified. In this study, the author investigated whether the 120 kDa protein of T. vaginalis may be involved in pathogenicity of trichomonads. Antibodies against 120 kDa protein of T. vaginalis, which was identified as pyruvate:ferredoxin oxidoreductase (PFOR) by peptide analysis of MALDI-TOF-MS, were prepared in rabbits. Pretreatment of T. vaginalis with anti-120 kDa Ab decreased the proliferation and adherence to vaginal epithelial cells (MS74) of T. vaginalis. Subcutaneous tissue abscess in anti-120 kDa Ab-treated T. vaginalis-injected mice was smaller in size than that of untreated T. vaginalis-infected mice. Collectively, the 120 kDa protein expressed by iron may be involved in proliferation, adhesion to host cells, and abscess formation, thereby may influence on the pathogenicity of T. vaginalis.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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• pp.74.1-74.1
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• 2007

See Full Text

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.563-569
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• 2004

The DNA sequence of the chitosanase gene (choK) from $\beta$-Proteobacterium KNU3 showed an 1,158-bp open reading frame that encodes a protein of 386 amino acids with a novel 74 signal peptide. The degenerated primers based on the partial deduced amino acid sequences from MALDI- TOF MS analyses yielded the 820 bp of the PCR product. Based on this information, double inverse PCR cloning experiments, which use the two specific sets of PCR primers rather than single set primers, identified the unknown 1.2 kb of the choK gene. Subsequently, a 1.8 kb of full choK gene was cloned from another PCR cloning experiment and it was then subcloned into pGEM T-easy and pUC18 vectors. The recombinant E. coli clone harboring recombinant pUC18 vector produced a clear halo around the colony in the glycol chitosan plates. The recombinant ChoK protein was secreted into medium in a mature form while the intracellular ChoK was produced without signal peptide cleavage. The activity staining of PAGE showed that the recombinant ChoK protein was identical to the chitosanase of wild-type. The comparison of deduced amino acid sequences of choK revealed that there is 92% identity with that of Sphingobacterium multivorum chitosanase. Judging from the conserved module in other bacterial chitosanases, chitosanase of KNU3 strain (ChoK) belongs to the family 80 of glycoside hydrolases.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.307-312
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• 2007

The present study was carried out to investigate differentially expressed chicken muscle proteins using proteomics approach. More than 300 protein spots were investigated for the muscle samples in 2DE gels and the differentially expressed protein spots between pectoralis and peroneus longus muscles from Cornish and White Leghorn breeds were characterized by MALDI-TOF. In pectoralis muscles, PGAM1 protein was detected as differentially expressed between White Leghorn and Cornish breeds. On the other hand, 4 protein spots (SP22, nxf-2, SOD1, TNNI2) were differentially expressed between White Leghorn and Cornish breeds in peroneus longus muscles. These proteins assumed to be related with muscle development, growth, stress, and movements in chicken. In this experimental process, 2D reference map of the chicken muscle proteins was needed and 25 proteins, which were commonly expressed in both pectoralis and peroneus longus muscles in both breeds, were selected and characterized. Upon finishing the exact roles of the differentially expressed proteins, the identified 5 proteins will be used as valuable information for the fundamental mechanisms of muscle biology and underline genetics.

• 한국수정란이식학회지
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• 제25권4호
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• Reproductive and Developmental Biology
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• 제33권3호
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• pp.183-187
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• 2009

This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is $2{\sim}4\;mm$ and $6{\sim}10\;mm$, were collected from ovary of slaughtered pig that each follicle of diameter $1{\sim}4\;mm$ and $6{\sim}10\;mm$. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $200{\mu}l$. Immobilized pH gradient(IPG) strip used 18 cm, $3{\sim}10\;NL$. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of $6{\sim}10\;mm$ follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.