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Chemical biology-based approaches on fluorescent labeling of proteins in live cells
Jung, Deokho,Min, Kyoungmi,Jung, Juyeon,Jang, Wonhee,Kwon, Youngeun The Royal Society of Chemistry 2013 Molecular bioSystems Vol.9 No.5
<P>Recently, significant advances have been made in live cell imaging owing to the rapid development of selective labeling of proteins <I>in vivo</I>. Green fluorescent protein (GFP) was the first example of fluorescent reporters genetically introduced to protein of interest (POI). While GFP and various types of engineered fluorescent proteins (FPs) have been actively used for live cell imaging for many years, the size and the limited windows of fluorescent spectra of GFP and its variants set limits on possible applications. In order to complement FP-based labeling methods, alternative approaches that allow incorporation of synthetic fluorescent probes to target POIs were developed. Synthetic fluorescent probes are smaller than fluorescent proteins, often have improved photochemical properties, and offer a larger variety of colors. These synthetic probes can be introduced to POIs selectively by numerous approaches that can be largely categorized into chemical recognition-based labeling, which utilizes metal-chelating peptide tags and fluorophore-carrying metal complexes, and biological recognition-based labeling, such as (1) specific non-covalent binding between an enzyme tag and its fluorophore-carrying substrate, (2) self-modification of protein tags using substrate variants conjugated to fluorophores, (3) enzymatic reaction to generate a covalent binding between a small molecule substrate and a peptide tag, and (4) split-intein-based C-terminal labeling of target proteins. The chemical recognition-based labeling reaction often suffers from compromised selectivity of metal–ligand interaction in the cytosolic environment, consequently producing high background signals. Use of protein–substrate interactions or enzyme-mediated reactions generally shows improved specificity but each method has its limitations. Some examples are the presence of large linker protein, restriction on the choice of introducible probes due to the substrate specificity of enzymes, and competitive reaction mediated by an endogenous analogue of the introduced protein tag. These limitations have been addressed, in part, by the split-intein-based labeling approach, which introduces fluorescent probes with a minimal size (∼4 amino acids) peptide tag. In this review, the advantages and the limitations of each labeling method are discussed.</P> <P>Graphic Abstract</P><P>This review summarizes various approaches to incorporate synthetic fluorescent probes to target proteins in live cells as well as current efforts to control the fluorescent signal using external stimuli. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c2mb25422k'> </P>
Photo-triggered fluorescent labelling of recombinant proteins in live cells
Jung, Deokho,Sato, Kohei,Min, Kyoungmi,Shigenaga, Akira,Jung, Juyeon,Otaka, Akira,Kwon, Youngeun The Royal Society of Chemistry 2015 Chemical communications Vol.51 No.47
<P>A method to photo-chemically trigger fluorescent labelling of proteins in live cells is developed. The approach is based on photo-caged split-intein mediated conditional protein trans-splicing reaction and enabled background-free fluorescent labelling of target proteins with the necessary spatiotemporal control.</P> <P>Graphic Abstract</P><P>A method to photo-chemically trigger fluorescent labelling of proteins in live cells is developed for background-free fluorescent labelling of target proteins with the necessary spatiotemporal control. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c5cc01067e'> </P>
AP 콜라겐 효소분해 펩타이드 섭취가 피부 주름, 탄력 및 보습 개선에 미치는 효과
정경미(Kyoungmi Jung),양미숙(Mi-Suk Yang),김승훈(Seung Hun Kim),노종화(Jonghwa Roh),김완기(Wangi Kim) 한국식품영양과학회 2021 한국식품영양과학회지 Vol.50 No.5
본 연구에서는 40~60세 성인 여성을 대상으로 APCP의 피부 주름, 탄력 및 보습 개선에 대한 이중눈가림, 무작위배정, 대조식품 비교 인체적용시험을 진행하였다. 눈가 주름 부위의 육안평가 및 Visiometer를 이용한 피부 주름 개선 평가에서 APCP 1,000 mg 및 1,500 mg 섭취군 모두 12주에 대조식품 섭취군 대비 유의적인 개선 효과를 보였고, 섭취종료 후 2주까지 피부 주름 개선이 유지됨을 확인할 수 있었다. 피부 탄력 개선 평가에서 R2, R5 및 R7 지표에 대해 APCP 1,000 mg 및 1,500 mg 섭취군 모두 6주부터 대조식품 섭취군 대비 유의적인 개선 효과를 보였고 12주까지 통계적으로 유의한 차이를 나타내었다. 피부 보습 개선 평가에서도 APCP 1,000 mg 및 1,500 mg 섭취군 모두 12주에 대조식품 섭취군 대비 유의적인 개선 효과를 보였고 섭취종료 후 2주까지 피부 보습 개선 상태가 유지되었다. 연구를 진행하는 동안 이상반응, 임상실험실 검사 및 활력징후 검사에서 유의미한 변화는 나타나지 않아 APCP의 안전성을 확인할 수 있었다. 본 인체적용시험 결과를 통해 APCP는 피부주름, 거칠기, 탄력 및 보습과 같은 피부 건강 개선에 도움을 줄 수 있는 건강기능식품임을 확인하였다. Collagen is a component of the human body, known to have a beneficial effect on human health when consumed as a food or dietary ingredient. This double-blinded, randomized, placebo-controlled study aimed to evaluate the efficacy and safety of AP collagen peptides (APCP) containing 3% glycine-proline-hydroxyproline, on improving skin wrinkles, elasticity and hydration. Totally, 105 subjects with crow’s-feet wrinkles were randomly assigned to receive either placebo, APCP 1,000 mg or APCP 1,500 mg, once daily for 12 weeks. Efficacy endpoints of skin wrinkles, elasticity and hydration were measured at baseline, 6 and 12 weeks, and at the 2-week follow-up visit. Compared to the placebo group, the visual evaluations and R3 (average roughness) values of crow’s-feet wrinkles were significantly decreased in both APCP groups at 12 weeks, and these effects were maintained until 2 weeks after completion of ingestion. During the intake period at 6 and 12 weeks, three parameters for skin elasticity (R2, R5, R7) showed significant improvement in both APCP groups, as compared to the placebo group. The APCP groups also showed a statistically significant difference in skin hydration compared to the placebo group, at 12 weeks and at 2 weeks after the end of intake. No adverse effects were encountered during the study period. These results indicate that APCP 1,000 mg and 1,500 mg can be used as a functional food ingredient to improve human skin conditions related to skin aging and hydration.
Kyoungmi Min,Suin Chae,Deokho Jung,권영은 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.5
Interfacial reactions kinetics often differ from kinetics of bulk reactions. Here, we describe how the density change of an immobilized reactant influences the kinetics of interfacial reactions. Self-assembled monolayers (SAMs) of alkanethiolates on gold were used as a model interface and the Diels-Alder reaction between immobilized quinones and soluble cyclopentadiene was used as a model reaction. The kinetic behavior was studied using varying concentrations of quinones. An unusual threshold density of quinones (Γ_c = 5.2-7.2%),at which the pseudo-first order rate constant started to vary as the reaction progressed, was observed. This unexpected kinetic behavior was attributed to the phase-separation phenomena of multi-component SAMs. Additional experiments using more phase-separated two-component SAMs supported this explanation by revealing a significant decrease in Γ_c values. When the background hydroxyl group was replaced with carboxylic or phosphoric acid groups, Γ_c was observed at below 1%. Also, more phase-separated thermodynamically controlled SAMs produced a lower critical density (3% < Γ_c < 4.9%) than that of the less phaseseparated kinetically controlled SAMs (6.5% < Γ_c < 8.9%).
강재헌,Nam Hyun-Jin,Jung Kyoungmi,Choi Gyeyoung,Lee Ji-Hae,Jeong Hyun Woo,Roh Jonghwa,Kim Wan-Gi 건강기능식품미래포럼 2021 건강기능식품미래포럼 학술지 Vol.1 No.3
Increasingly westernized eating habits and insufficient physical activity have increased prevalence of type 2 diabetes in Korea. Green tea extracts (GTE) obtained from Camellia sinensis have shown beneficial effects particularly on glucose metabolism and transport. Thus, many studies have been done on GTE for its actions on glucose control and insulin sensitivity. However, most of them were performed on cells and animals and not so many were on humans. Therefore, we conducted an open-label, crossover clinical study using AP GTE provided by AmorePacific Corporation R&D Center and investigated its effects on postprandial blood glucose after a high-fat/high-carbohydrate meal. Twenty subjects (men and women) aged ≥ 19 years with a body mass index (BMI) of 18.5–29.9 kg/m2 and fasting glucose of 100–139 mg/dL were enrolled. First, the control study was performed, where all subjects were fasted for 12 hours and blood (35 mL) was collected before (0 hours) they consumed a high-fat/high-carbohydrate meal and at 0.5, 1, 2, 3, 4, and 5 hours after the meal. After at least a week of washout period, AP GTE study was performed in the same way but taking 4 tablets of AP GTE after ingesting the same high-fat/high-carbohydrate meal. The blood samples were used for assessment of glucose and insulin concentrations. AP GTE-treated subjects had significantly lower plasma concentrations of glucose and insulin over 5 hours than the subjects not treated with AP GTE. During the experiment, no adverse effects were observed from the clinical laboratory testing, vital signs, and physical examinations. These results suggest that AP GTE supplementation may give beneficial effects to subjects who need glycemic control.
Progress and plans for the physics module development at KIAPS
Emilia Kyung Jin,Gill-Ran Jeong,Soo Ya Bae,Jung Hyo Chae,Hyun-Joo Choi,In Jin Choi,Ji-Young Han,Jung-Yoon Kang,So-Young Kim,Taekyun Kim,Joonsuk Lee,Kyung Hee Seol,Kyoungmi Cho,Juwon Lee,Dongil Jang,Hy 한국기상학회 2013 한국기상학회 학술대회 논문집 Vol.2013 No.4