• Interdisciplinary Bio Central
• /
• 제1권1호
• /
• pp.3.1-3.6
• /
• 2009

Introduction: GTPases known as translation factor play a vital role as ribosomal subunit assembly chaperone. The bacterial Obg proteins ($Spo{\underline{0B}}$-associated ${\underline{G}}TP$-binding protein) belong to the subfamily of P-loop GTPase proteins and now it is considered as one of the new target for antibacterial drug. The majority of bacterial Obgs have been commonly found to be associated with ribosome, implying that these proteins may play a fundamental role in ribosome assembly or maturation. In addition, one of the experimental evidences suggested that Bacillus subtilis Obg (BsObg) protein binds to the L13 ribosomal protein (BsL13) which is known to be one of the early assembly proteins of the 50S ribosomal subunit in Escherichia coli. In order to investigate binding mode between the BsObg and the BsL13, protein-protein docking simulation was carried out after generating 3D structure of the BsL13 structure using homology modeling method. Materials and Methods: Homology model structure of BsL13 was generated using the EcL13 crystal structure as a template. Protein-protein docking of BsObg protein with ribosomal protein BsL13 was performed by DOT, a macro-molecular docking software, in order to predict a reasonable binding mode. The solvated energy minimization calculation of the docked conformation was carried out to refine the structure. Results and Discussion: The possible binding conformation of BsL13 along with activated Obg fold in BsObg was predicted by computational docking study. The final structure is obtained from the solvated energy minimization. From the analysis, three important H-bond interactions between the Obg fold and the L13 were detected: Obg:Tyr27-L13:Glu32, Obg:Asn76-L13:Glu139, and Obg:Ala136-L13:Glu142. The interaction between the BsObg and BsL13 structures were also analyzed by electrostatic potential calculations to examine the interface surfaces. From the results, the key residues for hydrogen bonding and hydrophobic interaction between the two proteins were predicted. Conclusion and Prospects: In this study, we have focused on the binding mode of the BsObg protein with the ribosomal BsL13 protein. The interaction between the activated Obg and target protein was investigated with protein-protein docking calculations. The binding pattern can be further used as a base for structure-based drug design to find a novel antibacterial drug.

• 대한소아치과학회지
• /
• 제43권1호
• /
• pp.109-116
• /
• 2016

오늘날 소아치과 진료에 있어서 진정법은 필수 불가결한 요소로 받아들여지고 있다. 이에 대한소아치과학회는 학회원들의 보다 안전하고 효율적인 진정법 시행을 돕기 위해 진정법 위원회를 결성하고 2014년 소아치과 개원의들의 진정법 사용에 대한 실태조사를 실시 한 바 있다. 이 설문조사 결과, 진정법을 시행하는 교육 코스가 만들어 진다면 참여할 의사가 있는지에 대한 질문에 응답자의 86%가 참여의사를 밝혔으며 진정법 교육과정에서 배우고 싶은 내용으로 안전한 약물용량과 가이드라인이 2위를 차지하여 1위인 응급상황 시 대처법 다음으로 교육받고 싶은 내용으로 나타났다. 그러나 약물을 선택하고 용량을 결정하기 위해서는 여러 가지 요소를 고려해야 하므로 용량을 일률적으로 정하여 지침을 제정하는 것은 현실적으로 어려운 일이고 실제적으로도 그 어떤 진정법 지침에도 약물의 용량을 제시해 놓지는 않았다. 성공적인 진정법을 위해 소아치과의사는 각 과정마다 개별적인 치료 계획을 수립해야 하며, 각 진정제의 고유의 약효뿐 만 아니라 다른 약제들 간의 상호 작용에 대하여 충분히 숙지하는 것이 중요하다. 이 종설에서는 문헌 고찰을 통해 소아치과 임상에서 자주 사용되는 진정제들에 대한 정보를 되새기고 의사용 처방편람(Physicians' Desk Reference: PDR) 및 소아치과 분야의 다양한 교과서의 추천용량을 정리하여 소개하고자 한다.

• Molecules and Cells
• /
• 제43권11호
• /
• pp.909-920
• /
• 2020

Cytosolic Ca²⁺ levels ([Ca²⁺]_c) change dynamically in response to inducers, repressors, and physiological conditions, and aberrant [Ca²⁺]_c concentration regulation is associated with cancer, heart failure, and diabetes. Therefore, [Ca²⁺]_c is considered as a good indicator of physiological and pathological cellular responses, and is a crucial biomarker for drug discovery. A genetically encoded calcium indicator (GECI) was recently developed to measure [Ca²⁺]_c in single cells and animal models. GECI have some advantages over chemically synthesized indicators, although they also have some drawbacks such as poor signal-to-noise ratio (SNR), low positive signal, delayed response, artifactual responses due to protein overexpression, and expensive detection equipment. Here, we developed an indicator based on interactions between Ca²⁺-loaded calmodulin and target proteins, and generated an innovative GECI sensor using split nano-luciferase (Nluc) fragments to detect changes in [Ca²⁺]_c. Stimulation-dependent luciferase activities were optimized by combining large and small subunits of Nluc binary technology (NanoBiT, LgBiT:SmBiT) fusion proteins and regulating the receptor expression levels. We constructed the binary [Ca²⁺]_c sensors using a multicistronic expression system in a single vector linked via the internal ribosome entry site (IRES), and examined the detection efficiencies. Promoter optimization studies indicated that promoter-dependent protein expression levels were crucial to optimize SNR and sensitivity. This novel [Ca²⁺]_c assay has high SNR and sensitivity, is easy to use, suitable for high-throughput assays, and may be useful to detect [Ca²⁺]_c in single cells and animal models.

• 대한한방부인과학회지
• /
• 제24권3호
• /
• pp.28-47
• /
• 2011

Objectives: This study was performed to investigate the effects of Angelicae gigantis Radix (AGR) which is one of the most useful herbal-drug to treat patients with Polycystic Ovary Syndrome (PCOS) in Oriental medicine on gene expression of ovary tissue. Methods: The effects of AGR on gene expression of ovary tissue resected from PCOS induced rats using single injection of $\ss$-Estradiol 17-valerate (EV) was measured using microarray technique, and the functional analysis on these genes was conducted. Results: Total 2,812 genes were up-regulated or down-regulated, 1,421 genes were up-regulated, 1,391 genes were down-regulated by induction of PCOS. Up-regulated genes were mainly involved in biological function such as cell signalling pathways and inflammatory response. Expression levels of 1,442 genes were restored to those of naive animals by administration of AGR. 558 genes were restored to those of naive animals, which were lowered by induction of PCOS. 884 genes were lowered to naive levels, which were elevated by induction of PCOS. The functions of restored genes were partially involved in the restoration of expression levels, which were changed by induction of PCOS. Especially, up-regulated gene by induction of PCOS were mainly involved in these changes. These results mean restorative effects of AGR on damaged functions by induction of PCOS. The network of total protein interactions was measured using cytoscape program, and some key molecules, such as IRS2, MCM10, ORC2L related in up-regulated genes, CTBP2, CD44, RHOA, related in down-regulated genes that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conclusion: Restored genes by AGR were thought to have common pathways related in regulation of gene expressions. Especially, genes in restored expression levels by AGR, which were up-regulated by induction of PCOS, were regulated by 9 of common transcription factors, genes in restored expression levels by AGR, which were down-regulated by induction of PCOS, were involved in 25 of common transcription factors.

• 생명과학회지
• /
• 제28권4호
• /
• pp.488-495
• /
• 2018

p53 유전자는 스트레스, DNA 손상, 저산소증 및 종양 발생에 대한 세포 반응의 전사 조절에서 중요한 역할을 담당한다. 최근에 발견된 다양한 종류의 p53의 생리 활성을 생각한다면 p53이 암 조절에 관여한다는 것은 놀랄만한 일이 아니다. 인간 암의 약 50%에는 p53 유전자의 돌연변이 또는 p53을 활성화시키는 기전의 결함을 통해 p53 단백질 기능의 불활성화가 나타난다. p53 기능의 이러한 장애는 p53 의존 반응으로부터 회피를 허용함으로써 종양의 진화에 결정적인 역할을 하게 된다. 최근의 많은 연구들은 p53의 돌연변이를 대폭 감소시키거나 p53의 종양 억제 기능을 복원하기 위하여 선택적인 저분자 화합물을 동정함으로써 p53의 돌연변이를 직접 표적하는 것에 초점을 두고 있다. 이들 저분자는 좋은 약물과 유사한 특성을 유지하면서 다양한 상호작용을 효과적으로 조절해야 한다. 이 중, p53의 음성조절인자 핵심인 MDM2의 발견은 p53과 MDM2 간의 상호작용을 차단하는 새로운 저분자 억제제의 설계를 제공하였다. 저분자 화합물 중 일부는 개념 증명 연구에서 임상 시험으로 옮겨졌으며 향후 맞춤형 항암제가 추가될 전망이다. 본 리뷰에서는 야생형 p53과 돌연변이 p53의 구조적 및 기능적 중요성과 p53을 직접 표적하는 치료제 개발, p53과 MDM2 간의 상호작용을 억제하는 화합물에 대하여 검토하였다.

• Bulletin of the Korean Chemical Society
• /
• 제31권1호
• /
• pp.52-58
• /
• 2010

Human histamine H1 receptor (HHR1) is a G protein-coupled receptor and a primary target for antiallergic therapy. Here, the ligand-based three-dimensional pharmacophore models were built from a set of known HHR1 inverse agonists using HypoGen module of CATALYST software. All ten generated pharmacophore models consist of five essential features: hydrogen bond acceptor, ring aromatic, positive ionizable and two hydrophobic functions. Best model had a correlation coefficient of 0.854 for training set compounds and it was validated with an external test set with a high correlation value of 0.925. Using this model Maybridge database containing 60,000 compounds was screened for potential leads. A rigorous screening for drug-like compounds unveiled RH01692 and SPB00834, two novel molecules for HHR1 with good CATALYST fit and estimated activity values. The new lead molecules were docked into the active site of constructed HHR1 homology model based on recently crystallized squid rhodopsin as template. Both the hit compounds were found to have critical interactions with Glu177, Phe432 and other important amino acids. The interpretations of this study may effectively be deployed in designing of novel HHR1 inverse agonists.

• 약학회지
• /
• 제59권6호
• /
• pp.270-277
• /
• 2015

There is a lack of evidence on the impact of pharmacist interventions in diabetes care in South Korea. The aim of this study was to investigate the effects of pharmacist counseling on clinical and humanistic outcomes in patients with type 2 diabetes. An uncontrolled before-and-after study was conducted at the outpatient diabetes clinic in a teaching hospital in Gyeongnam, South Korea between January 2 and November 30, 2014. A total of 37 patients were enrolled. During the study periods, the participants and pharmacists met every 2 weeks via follow-up telephone calls in addition to face-to-face meetings upon returning for care visit. At each meeting, a structured patient counseling was performed and the average duration of each meeting was 15~30 minutes. The participants were requested to record daily self-care activities. At the end of this study, patients' satisfaction on pharmacist care was evaluated using the questionnaire developed by us. Compared to baseline, significant reductions (mean${\pm}$standard deviation, p<0.05) in HbA1c were observed at each follow-up period: $-0.32{\pm}0.72%$ from baseline to 3 months; $-0.52{\pm}0.76%$ from baseline to 6 months; $-0.72{\pm}0.76%$ from baseline to 9 months. Over the same follow-up period, the proportions of patients achieving target HbA1c (defined as HbA1c<6.5%) were 3.1%, 10.3%, and 20%, respectively. The proportions of patients who never missed a dose during the same follow-up period were 43.8%, 31.0%, and 20.0%, respectively. The results from the patient satisfaction survey indicated that pharmacist counseling improved patients' knowledge about diabetes and possible drug interactions. Especially, the information on a healthy diet and lifestyle was the most satisfying. To conclude, follow-up telephone calls combined with face-to-face meetings improved clinical and humanistic outcomes in patients with type 2 diabetes mellitus.

• Journal of Microbiology and Biotechnology
• /
• 제22권12호
• /
• pp.1659-1664
• /
• 2012

TSAHC [4'-(p-toluenesulfonylamido)-4-hydroxychalcone] is a promising antitumorigenic chalcone compound, especially against TM4SF5 (four-transmembrane L6 family member 5)-mediated hepatocarcinoma. We evaluated the potential of TSAHC to inhibit the catalytic activities of nine cytochrome P450 isoforms and of P-glycoprotein (P-gp). The abilities of TSAHC to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The P-gp inhibitory effect of TSAHC was assessed by [$^3H$]digoxin accumulation in the LLCPK1-MDR1 cell system. TSAHC strongly inhibited CYP2C8, CYP2C9, and CYP2C19 isoform activities with $K_i$ values of 0.81, 0.076, and $3.45{\mu}M$, respectively. It also enhanced digoxin accumulation in a dose-dependent manner in the LLCPK1-MDR1 cells. These findings indicate that TSAHC has the potential to inhibit CYP2C isoforms and P-gp activities in vitro. TSAHC might be used as a nonspecific inhibitor of CYP2C isoforms based on its negligible inhibitory effect on other P450 isoforms such as CYP1A2, CYP2A6, CYP2B6, CYP2D6, CYP2E1, and CYP3A.

• 한국자기공명학회논문지
• /
• 제12권1호
• /
• pp.1-13
• /
• 2008

Acetyl-CoA carboxylase (ACC) catalyzes the first step in fatty acid biosynthesis: the synthesis of malonyl-CoA from acetyl-CoA. As essential regulators of fatty acid biosynthesis and metabolism, ACCs are regarded as therapeutic targets for the treatment of metabolic diseases such as obesity, In ACC, the biotinoyl domain performs a critical function by transferring an activated carboxyl group from the biotin carboxylase domain to the carboxyl transferase domain, followed by carboxyl transfer to malonyl-CoA. Despite the intensive research on this enzyme, only the bacterial and yeast ACC structures are currently available, To explore the mechanism of ACC holoenzyme function, we determined the structure of the biotinoyl domain of human ACC2 and analyze its characteristics using NMR spectroscopy. The 3D structure of the hACC2 biotinoyl domain has a similar folding topology to the previously determined domains from E. coli and P. Shermanii, however, the 'thumb' structure is absent in the hACC2 biotinoyl domain. Observations of the NMR signals upon the biotinylation indicate that the biotin group of hACC2 does not affect the structure of the biotinoyl domain, while the biotin group for E. coli ACC interacts directly with the thumb residues that are not present in the hACC2 structure. These results imply that, in the E. coli ACC reaction, the biotin moiety carrying the carboxyl group from BC to CT can pause at the thumb of the BCCP domain. The human biotinoyl domain, however, lacks the thumb structure and does not have additional non-covalent interactions with the biotin moiety; thus, the flexible motion of the biotinylated lysine residue must underlie the "swinging arm" motion. This study provides insight into the mechanism of ACC holoenzyme function and supports the "swinging arm" model in human ACCs.

• 생명과학회지
• /
• 제17권3호통권83호
• /
• pp.334-339
• /
• 2007

본 연구는 우황청심원을 비롯한 상용되는 20종의 한약제제를 대상으로 9종의 시토크롬 동종효소에 대한 대사능의 저해정도를 고속 스크리닝 기법을 이용하여 탐색함으로써, 한약제제와 약물의 병용으로 인한 약물 상호작용 가능성을 평가하고자 하였다. 인체 간 마이크로좀 시료에 9종의 주요 시토크롬 약물대사효소의 지표약물과NADPH-generating system및 한약제제(500 ${\mu}g/ml$)를 첨가한 후 $37^{\circ}C$에서 15분간 반응시켜 생성된 각각의 대사물을 LC/MS/MS를 이용하여 정량하여 시토크롬 동종효소 활성의 변화를 평가하였다. 그 결과 우황청심원 현탁액 및 황련해독탕 물 추출물이 각각 CYP2B6 및 CYP2D6 효소 활성을 선택적으로 강력하게 저해하였다. 이러한 결과는 약국에서 쉽게 구입할 수 있는 한약제제들 중 일부는 인체 간 시토크롬 활성 저해능을 가지고 있고, 이들 효소에 의해 대사되는 약물과의 병용 복용시 약물상호작용 발생 가능성이 있음을 의미한다. 향후 한약제제에서 저해능을 나타내는 주된 성분을 규명하여 이 성분의 저해능과 저해 기전을 살피는 노력이 필요할 것이다.