• 제목/요약/키워드: glycerophosphate

검색결과 71건 처리시간 0.024초

대장균 트립토판 생성효소의 소단위체간 상호조절 (Intersubunit Communication of Escherichia coli Tryptophan Synthase)

  • 조원진;임운기
    • 생명과학회지
    • /
    • 제27권12호
    • /
    • pp.1410-1414
    • /
    • 2017
  • 대장균 트립토판 생성효소는 ${\alpha}_2{\beta}_2$ 복합체로 구성되며, 트립토판 생합성에서 최종 2 단계의 반응에 관여한다. 두 개의 소단위체는 분자 터널로 연결되어 있어, 기질 채널링이 일어난다. 활성 부위간 상호 조절하는 정교한 조절 기작에는 ${\alpha}$-루프 L6(${\alpha}L6$), ${\alpha}L2$, ${\alpha}L3$이 관여한다. 본 연구에서는 이 자리의 잔기치환체를 써서 소단위체에 특이적으로 결합하는 리간드의 영향을 조사하여 소단위체간 상호 조절기작에 따른 구조 변화를 살펴보았다. ${\alpha}TS$의 활성부위에 결합하는 D,L-${\alpha}$-glycerophosphate(GP)는 모든 잔기치환체를 야생형 수준으로 회복시켰다. ${\beta}TS$의 기질인 L-Ser는 다양한 효과를 나타낸다. 야생형과 NS104에서는 속도가 감소한 반면, GD51과 PH53에서는 거의 영향이 없었고, PT53와 DG56은 증가하였다. 이는 반응 중간 화학종의 분포의 변화와 연관될 가능성을 제시한다. GP와 L-Ser를 동시에 처리했을 때는 특이하게도 PH53는 가장 안정한 잔기치환체였다. 이는 Pro53가 소단위체간의 조절기작에서 중요한 역할을 하는 것을 시사한다.

온도감응성 키토산 중성용액을 이용한 약물송달시스템 (Temperature-Sensitive Drug Delivery System of Acetaminophen Using Neutral Chitosan Solution)

  • 김호정;이화정;구영순
    • Journal of Pharmaceutical Investigation
    • /
    • 제38권4호
    • /
    • pp.229-234
    • /
    • 2008
  • In the present study, chitosan-glycerophosphate sodium salt solution as a thermosensitive system (TSS) was used to formulate a temperature-sensitive drug delivery system (TSDDS) containing acetaminophen (AAP). The optimized TSS was prepared by measuring gelation temperature, gelation time and rheological properties of TSS. The optimized gelation temperature and time of TSS were $36^{\circ}C$ and 100 seconds, respectively. The viscosity of TSS was also suitable for maintaining gel structure at $37.2^{\circ}C$. The release profiles of TSDDS in PBS/pH 7.4 with various apparatuses and mass loss of TSDDS were investigated. The time required to release 50% of AAP from TSDDS ($t_{50%}$) was 120 min with the formation of pore on the surfaces, which was 2 times longer than that from AAP-chitosan gel. In addition, TSDDS was degraded approximately 80% within 4 hr and then degraded slowly for 20 hrs. In conclusion, AAP-TSS (TSDDS) formulated in this study might be suitable for some specific uses such as subcutaneous injection and rectal formulation.

Isozyme electrophoresis patterns of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China

  • Park, Gab-Man;Yong, Tai-Woon;Im, Kyung-Il;Lee, Kyu-Je
    • Parasites, Hosts and Diseases
    • /
    • 제38권1호
    • /
    • pp.45-48
    • /
    • 2000
  • An enzyme analysis of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China was conducted using a horizontal. starch gel electrophoresis in order to elucidate their genetic relationships. A total of eight enzymes was employed from two different kinds of buffer systems. Two loci from each enzyme of aconitase and esterase (${\alpha}-Na{\;}and{\;}{\beta}-Na$) : and only one locus each from six enzymes, gluucose-6-phosphate dehydrogenase (G6PD), ${\alpha}-glycerophosphate$ dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphoglucomutase (PGM) were detected. Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. However, esterase (${\alpha}-Na$), GPD, HBDH and PGI loci showed polymorphic banding patterns. Two populations of C. sinensis were more closely clustered within the range of genetic identity value of 0.998-1.0. In summarizing the above results, two populations of C. sinensis employed in this study showed mostly monomorphic enzyme protein banding patterns, and genetic differences specific between two populations.

  • PDF

덱사메타존이 골아유사세포의 성장과 분화에 미치는 영향 (The Effects of Dexamethasone on Growth and Differentiation of Osteoblast-like Cell)

  • 이재목
    • Journal of Periodontal and Implant Science
    • /
    • 제29권2호
    • /
    • pp.277-289
    • /
    • 1999
  • The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. To evaluate the effects of Dex growth and differentiation of MC3T3-E1 cells, cells were seeded in alpha-modified eagle medium containing 10% fetal bovine serum, 10mM beta-glycerophosphate , $50{\mu}g/ml$ of ascorbic acid, with or without $10^{-7}M$ Dex and examined cell proliferation activities, alkaline phosphatase activities, and bone nodule formation until 25days. The results were as follows : 1. In Dex group, cell proliferation activities were lower until 15 days compared to control group. Bone nodules formation were showed at 10 days. 2. In the time-response effect, ALP activities were increased until the 10 days in control groups thereafter decreased and ALP activities of Dex group were lower aspect than control group until the 10 days In this study, bone nodule formation of osteoblast-like cells were accelerated by Dex and cell proliferation activities, ALP activity of Dex group showed lower than control group. Dex was considered that it did suppress initial growth, but accerelate mineralization of osteoblast-like cells.

  • PDF

Effects of Culture Conditions on Osteogenic Differentiation in Human Mesenchymal Stem Cells

  • Song, Su-Jin;Jeon, O-Ju;Yang, Hee-Seok;Han, Dong-Keun;Kim, Byung-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권7호
    • /
    • pp.1113-1119
    • /
    • 2007
  • Human bone marrow-derived mesenchymal stem cells (hBMMSCs) must differentiate into osteogenic cells to allow for successful bone regeneration. In this study, we investigated the effects of different combinations of three soluble osteogenic differentiation-inducing factors [L-ascorbic acid (AC), ${\beta}$-glycerophosphate (${\beta}G$), and bone morphogenic protein-2 (BMP-2)] and the presence of a hydroxyapatite (HA) substrate on hBMMSC osteogenic differentiation in vitro. hBMMSCs were cultured in medium containing various combinations of the soluble factors on culture plates with or without HA coating. After 7 days of culture, alkaline phosphatase (ALP) activity, calcium deposition, and osteoprotegerin (OPG) and osteopontin (OPN) expression were measured. The effects of individual and combined factors were evaluated using a factorial analysis method. BMP-2 predominantly affected expression of early markers of osteogenic differentiation (ALP and OPG). HA had the highest positive effect on OPN expression and calcium deposition. The interaction between AC, ${\beta}G$, and HA had the second highest positive effect on ALP activity.

Osteogenic Differentiation Potential in Parthenogenetic Murine Embryonic Stem Cells

  • Kang, Ho-In;Cha, Eun-Sook;Choi, Young-Ju;Min, Byung-Moo;Roh, Sang-Ho
    • International Journal of Oral Biology
    • /
    • 제33권3호
    • /
    • pp.91-95
    • /
    • 2008
  • Embryonic stem cells have a pluripotency and a potential to differentiate to all type of cells. In our previous study, we have shown that embryonic stem cells (ESCs) lines can be generated from murine parthenogenetic embryos. This parthenogenetic ESCs line can be a useful stem cell source for tissue repair and regeneration. The defect in full-term development of parthenogenetic ESCs line enables researchers to avoid the ethical concerns related with ESCs research. In this study, we presented the results demonstrating that parthenogenetic ESCs can be induced into osteogenic cells by supplementing culture media with ascorbic acid and $\beta$-glycerophosphate. These cells showed morphologies of osteogenic cells and it was proven by Von Kossa staining and Alizarin Red staining. Expression of marker genes for osteogenic cells (osteopontin, osteonectin, alkaline phosphatase, osteocalcin, bone-sialoprotein, collagen type1, and Cbfa1) also confirmed osteogenic potential of these cells. These results demonstrate that osteogenic cells can be generated from parthenogenetic ESCs in vitro.

A Trial of Screening of Genes Involved in Odontoblasts Differentiation from Human Dental Pulp Stem Cells

  • Park, Yoon-Kyu;Kim, Hyun-Jin
    • International Journal of Oral Biology
    • /
    • 제37권4호
    • /
    • pp.167-173
    • /
    • 2012
  • This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

Purification and Characterization of a Thermostable Alkaline Phosphatase Produced by Thermus caldophilus GK24

  • Kim, You-Jin;Park, Tae-Shin;Kim, Hyun-Kyu;Kwon, Suk-Tae
    • BMB Reports
    • /
    • 제30권4호
    • /
    • pp.262-268
    • /
    • 1997
  • The thermophilic and thermostable alkaline phosphatase was purified to near homogeneity from the osmotic lysis of Thermus caldophilus GK24, The purified enzyme had an apparent molecular mass of 108, 000 Da and consisted of two subunits of 54,000 Da. lsoelectric-focusing analysis of the purified enzyme showed a pi of 7.3. The enzyme contained two Cys residues, and its amino acids composition was quite different from that of Thermus aquaticus YT-1 alkaline phosphatase and Escherichia coli alkaline phosphatase, The optimum pH and temperature of the enzyme were 11.0-11.5 and $80^{\circ}C$ respectively. The enzyme was stable in the pH range of 9.0-12.0 at $25^{\circ}C$ for 36 h. and the half-life at $80^{\circ}C$ (pH 11.0) was 6 h. The enzyme was activated by $MgCl_2$ and inhibited by EDTA. With ${\rho}-nitrophenyl\;phosphate\;({\rho}NPP)$ as the substrate, the enzyme had a Michaelis constant $(K_m) $of $3.6{\times}10^{-5}M$, The enzyme preferentially hydrolyzed the phosphomonoester bond of AMP in ribonucleotides and glycerophosphate.

  • PDF

Preparation and Drug-releasing Properties of Chitosan-based Thermosensitive Composite Hydrogel

  • Li, Heping;Yu, Tao;Li, Shan;Qin, Long;Ning, Jingheng
    • 대한화학회지
    • /
    • 제56권4호
    • /
    • pp.473-477
    • /
    • 2012
  • The novel chitosan-based thermosensitive hydrogels were prepared as control-releasing drug carriers. N-carboxyethyl chitosan (ACS) was synthesized by microwave heating for 1 h through Michael addition of CS to acrylic acid in a grafting yield of 52.97%, which was proved to be a faster and more efficient way than ordinary methods. 5-Fu was modified with formaldehyde to synthesize N,N'-Bis(hydroxymethyl)-5-fluorouracil (5-Fu-OH). Then an esterification was performed using ACS and 5-Fu-OH to give 5-Fu-ACS. The new thermosensitive hydrogels were prepared by adding sodium glycerophosphate to the solution of compounds under a certain constant temperature. Simultaneously, the hydrogels' swelling rate, in vitro drug release rate and thermosensitive were studied, and found that the 5-Fu-ACS composite hydrogel had more excellent releasing effect, higher drug loading and better thermosensitive.

다공성 티타늄 임플란트의 생체적합성 증진을 위한 복합 표면처리에 관한 연구 (A Study of Multi-Surface Treatments on the Porous Ti Implant for the Enhancement of Bioactivity)

  • 조유정;김영훈;장형순;강태주;이원희
    • 한국재료학회지
    • /
    • 제18권5호
    • /
    • pp.229-234
    • /
    • 2008
  • Porous Ti implant samples were fabricated by the sintering of spherical Ti powders in a high vacuum furnace. To increase their surface area and biocompatibility, anodic oxidation and a hydrothermal treatment were then applied. Electrolytes in a mixture of glycerophosphate and calcium acetate were used for the anodizing treatment. The resulting oxide layer was found to have precipitated in the phase form of anatase $TiO_2$ and nano-scaled hydroxyapatite on the porous Ti implant surface. The porous Ti implant can be modified via an anodic oxidation method and a hydrothermal treatment for the enhancement of the bioactivity, and current multi-surface treatments can be applied for use in a dental implant system.