• 한국미생물·생명공학회지
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• 제24권4호
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• pp.408-414
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• 1996

The SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane, and has been known to be rate-limiting factor of secretion in Escherichia coli. In order to study the extracellular protein secretion in Gram-positive microorganism, we have, constructed strains harboring more than one copy of the gene for SecY. Firstly, the gene, for B. subtilis SecY and its promoter region was subcloned into pDH32 and the chimeric vector was inserted into amyE locus by homologous recombination. Secondly, low copy number vector, pCED6, was also used for subcloning the secY gene and for constructing a strain which harbors several copies of secY. The KH1 cell which harbor two copies of secY on the chromosome excreted more extracellular proteins than the wild type PB2. Moreover, the KH2 cells which harbor several copies of secY in pCED6 vector excreted more extracellular proteins than the KH1 cells. Here, we found that the capacity of protein secretion is partly controlled by the number of secY and it is suggested that SecY has also an important role in protein secretion in B. subtilis, a gram positive microorganism, as like in E. coli. This will promote the use of B. subtilis as a host for the expression of useful foreign gene and excretion of precious proteins.

• 한국자원식물학회지
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• 제30권6호
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• pp.640-646
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• 2017

In this study, we elucidated anti-cancer activity and potential molecular mechanism of 70% ethanol extracts from Taxilli Ramulus (Taxillus chinensis (DC.) Danser) (TR-E70) against human colorectal cancer cells. Anti-cell proliferative effect of TR-E70 was evaluated by MTT assay. The effect of TR-E70 on the expression of cyclin D1 in the protein and mRNA level was evaluated by Western blot and RT-PCR, respectively. TR-E70 suppressed the proliferation of human colorectal cancer cell lines, HCT116 and SW480. Although TR-E70 decreased cyclin D1 expression in protein and mRNA level, decreased level of cyclin D1 protein by TR-E70 more dramatically occurred than that of cyclin D1 mRNA. Cyclin D1 downregulation by TR-E70 was attenuated in presence of MG132. In addition, TR-E70 phosphorylated threonine-286 (T286) of cyclin D1. TR-E70-mediated cyclin D1 degradation was blocked in presence of LiCl as an inhibitor $GSK3{\beta}$ but not PD98059 as an ERK1/2 inhibitor and SB203580 as a p38 inhibitor. Our results suggest that TR-E70 may downregulate cyclin D1 as one of the potential anti-cancer targets through $GSK3{\beta}$-dependent cyclin D1 degradation. From these findings, TR-E70 has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

• 한국어병학회지
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• 제17권3호
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• pp.191-198
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• 2004

$estradiol-17\beta$ ${E_2}$에 노출된 미성숙 조피볼락의 혈장 내 vitellogenin (VTG), alkaline-labile protein phosphorus (ALPP), calcium (Ca), glutamate pyruvate transaminase (GPT) 및 hepatosomatic index (HSI)의 일시적인 변화가 조사되었다. 70% 에탄올에 녹여진 ${E_2}$ (5 mg/kg B.W.)를 조피볼락 치어의 복강에 주사한 후 0, 1, 3, 6, 9, 12 및 15일에 혈장이 수집되었다. 전기 영동상에서 주사 후, 3일째 약 170 kDa의 위치에 염색성을 나타내는 VTG 밴드가 검출되었다. 밴드의 염색성은 6일째 더욱 강하게 나타났지만, 시간의 경과와 더불어 흐려져 15일째에는 노출 전 처럼 VTG 밴드가 관찰되지 않았다. 혈장 내 ALPP와 Ca는 1일째부터 급격히 증가했으며 VTG 변동과 유사하게 6일째 가장 높은 농도를 나타낸 후, 시간의 경과와 더불어 점점 감소해 15일째는 노출 전과 유사한 값을 나타내었다. GPT는 투여 1일부터 급격히 증가해 3일에 최고치를 나타낸 이후 시간의 경과와 더불어 감소하는 경향을 나타냈으며 HSI의 경우도 1일째부터 증가해 3일째에 가장 높은 수치를 나타낸 후, 점점 감소해 ${E_2}$ 투여전과 유사한 수치를 나타내었다. 이들 결과로 E2에 노출 된 이후 조피볼락의 혈장 내 ALPP, Ca, GPT 및 HSI는 혈장 VTG와 유사한 증․감을 나타내는 것을 알 수 있었다. 따라서, ALPP, Ca, GPT 및 HSI는 VTG와 더불어 연안 생태계 내에서 외인성 ${E_2}$ 노출에 대한 생물학적 지표로서 사용 가능할 것으로 판단되어진다.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.934-944
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• 2007

Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

• The Journal of Korean Physical Therapy
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• 제19권4호
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• pp.1-14
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• 2007

Background: It is generally accepted that skeletal muscle contraction is triggered by nerve impulse and intracellular $Ca^{2+}\;([Ca^{2+}]_i)$ released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR). Specifically, this process, called excitation-contraction (E-C) coupling, takes place at intracellular junctions between the plasma membrane, the transverse (T) tubule L-type $Ca^{2+}$ channel (dihydropyridine-sensitive L-rype $Ca^{2+}$ channel, DHPR, also called tetrads), and the SR $Ca^{2+}$ release channel (ryanodine-sensitive $Ca^{2+}$ release channel, RyR, also called feet) of internal $Ca^{2+}$ stores in skeletal muscle cells. Furthermore, it has been reported that the $Ca^{2+-}$ dependent and -independent contraction determine the expression of skeletal muscle genes, thus providing a mechanism for tightly coupling the extent of muscle contraction to regulation of muscle plasticity-related excitation-transcription (E-T) coupling. Purpose: Expression and activity of plasticity-associated enzymes in gastrocnemius muscle strips have not been well studied, however. Methods: Therefore, in this study the expression and phosphorylation of E-C and E-T coupling-related mediators such as protein kinases, ROS(reactive oxygen species)- and apoptosis-related substances, and others in gastrocnemius muscles from rats was examined. Results: I found that expression and activity of MAPKs (mitogen-activated protein kinases, ERK1/2, p38MAPK, and SAPK/JNK), apoptotic proteins (cleaved caspase-3, cytochrome c, Ref-1, Bad), small GTP-binding proteins (RhoA and Cdc42), actin-binding protein (cofilin), PKC (protein kinase C) and $Ca^{2+}$ channel (transient receptor potential channel 6, TRPC6) was observed in rat gastrocnemius muscle strips. Conclusion: These results suggest that MAPKs, ROS- and apoptosis-related enzymes, cytoskeleton-regulated proteins, and $Ca^{2+}$ channel may in part functionally import in E-C and E-T coupling from rat skeletal muscles.

• 약학회지
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• 제48권6호
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• pp.328-334
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• 2004

Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

• BMB Reports
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• 제45권1호
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• pp.14-19
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• 2012

A feruloyl esterase encoding gene (designated fae6), derived from a leachate metagenomic library, was cloned and the nucleotide sequence of the insert DNA determined. Translational analysis revealed that fae6 consists of a 515 amino acid poly-peptide, encoding a 55 kDa pre-protein. The Fae6 primary structure contained the G-E-S-A-G sequence, which corresponds well with a typical catalytic serine sequence motif (G-x-S-x-G). The fae6 gene was successfully over-expressed in E. coli and the recombinant protein was purified to 8.4 fold enrichment with 17% recovery. The $K_M$ data showed Fae6 has a high affinity to methyl sinapate while thermostability data indicated that fae6 was thermolabile with a half life ($T_{1/2}$) < 30 min at $50^{\circ}C$. High affinity for Fae6 against methyl sinapate, methyl ferulate and ethyl ferulate suggest that the enzyme can be useful in hydrolyzing ferulated polysaccharides in a biorefinery process.

• 대한지역사회영양학회지
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• 제10권6호
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• pp.880-891
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• 2005

The purpose of this study was to investigate nutrients intakes and nutritional quality of Adults and Elderly People in an island area (Wando). A three-day dietary intakes survey, using a 24 hour recall method was obtained from 187 subjects aged 46 to 84 (mean age 65.3) living in an island area (Wando). Nutient intakes were analyzed using CAN-Pro soft program and compared to Korean RDA. The quality of nutrients was assessed by analyzing nutrient adequacy ratio (NAR), mean adequacy ratio (MAR), and index of nutritional quality (INQ). The average daily mean energy intakes were 1869.0 kcal for males and 1943.9 kcal for females, respectively. Daily intakes of protein for males and females were 28.0 and 30.4 g and those of fat were 31.5 and 28.51 g, respectively. Nutrient consumed below $75\%$ of Korean RDA was protein, vitamin A, Ca and Zn in both males and females. Average CPF ratio of males and females were 78.8 : 6.0 : 15.1 and 80.0 : 6.4 13.4, respectively. Energy intake ratio from protein was significantly higher in over 60 years males. Carbohydrate dependency decreased with age. Protein dependency increased with age. Nutrient adequacy ratio (NAR) of energy, protein vitamin A and vitamin E were increased with age in males. The mean adequacy ratio (MAR), an index of overall dietary quality, were not significantly different by age group. Average MAR for males and females was 0.77 and 0.78, respectively. The index of nutritional quality (INQ) showed the tendency to decrease with age. Especially, there were significantly decreases in INQ of all nutrients, except protein, with age. Based on these results, it is evident that people in the island area did not consume enough nutrients. Specially, dietary intake of protein, vitamin A, vitamin E, and Ca were not adequate. (Korean J Community Nutrition 10(6) : $880\∼891$, 2005)

• 미생물학회지
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• 제30권6호
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• pp.421-424
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• 1992

Bacillus megaterium ATCC14945의 페니실린 지 아실라제 유전자를 갖는 Escherichia coli JM83이 한 종류의 단백질을 대량으로 만들어 내었다(전체 세포 단백질 양의 20%이상). 이 단백질은 아미노 말단의 아미노산 서열 분석을 통해서 E. coli heat protein인 GroEL로 확인되었다. 또한 이 groEL 단백질은 외래의 페니실린 지 아실라제가 발현됨으로써 27과 37도 두 온도에서 생산됨을 알았다.

• 한국콘텐츠학회논문지
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• 제8권6호
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• pp.74-81
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• 2008

단백질 상호작용 네트워크는 허브(hub)라 할 수 있는 상호작용 수가 많은 소수의 단백질과 상호작용수가 적은 다수의 단백질들로 구성된다. 최근 들어 여러 연구들에서 허브 단백질이 비 허브(non-hub) 단백질보다 상호작용 네트워크에 필수적인 단백질일 가능성이 높다고 보고되고 있다. 이러한 현상을 중심-치명 룰(centrality-lethality rule)이라 하는데, 이는 복잡계 네트워크에서 허브단백질의 중요성 및 네트워크 구조의 중요성을 설명하기 위한 방법으로 폭넓게 신뢰받고 있다. 이에 본 논문에서는 중심-치명 룰이 항상 옳게 적용되는지를 확인하기 위해 Uetz, Ito, MIPS, DIP, SGD, BioGRID와 같은 효모에 관한 공개된 모든 단백질 상호작용 데이터베이스들을 분석하였다. 흥미롭게도, 상호작용 데이터가 적은 데이터베이스들(Uetz, Ito, DIP)에서는 중심-치명 룰을 잘 나타냈지만 상호작용 데이터가 대용량인 데이터 베이스들(SGD, BioGRID)에서는 중심-치명 룰이 잘 맞지 않음을 확인하였다. 이에 따라 SGD와 BioGRID 데이터베이스로 부터 얻은 상호작용 네트워크의 특징을 분석하고 DIP 데이터베이스의 상호작용 네트워크와 비교하였다.