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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal DOI :
Korean Society for Biochemistry and Molecular Biology
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Volume & Issues
Volume 40, Issue 6 - Nov 2007
Volume 40, Issue 5 - Sep 2007
Volume 40, Issue 4 - Jul 2007
Volume 40, Issue 3 - May 2007
Volume 40, Issue 2 - Mar 2007
Volume 40, Issue 1 - Jan 2007
Selecting the target year
The Effect of Benzene on the Activity of Adenosine Deaminase in Tissues of Rats
Turhan, Ali ; Dere, Egemen ;
BMB Reports , volume 40, issue 3, 2007, Pages 295~301
DOI : 10.5483/BMBRep.2007.40.3.295
Swiss Albino (Rat rattus norvegicus) rats were intraperitoneally injected with a 100 mg
dosage of benzene, a toxic and carcinogenic agent widely used for industrial purposes. Changes in the adenosine deaminase (ADA) activity in the liver, kidney and serum of rats were investigated at 0, 2, 4, 8, 16, 32 and 64 h following injection. Serum physiological was administered to each control group. Enzyme activities were measured spectrophotometrically. Our purpose was to further investigations of some diseases caused by benzene, and present evidence of variations in the activity of ADA enzyme effected by benzene. While benzene caused significant inhibitions in ADA activity in the liver at 16 and 32 h and at 0.05 probability level, no significant inhibition or activation occurred at other test periods (hours). ADA activity did not present any significant variation in the kidneys. It was observed that ADA activity displayed similar patterns in the control groups. Comparisons of ADA activities in the two groups showed a statistically significant decrease between
hours (p< 0.05), demonstrating a direct correlation between benzene and its effects on ADA enzymes.
A Proteomic Screen for Presynaptic Terminal N-type Calcium Channel (CaV2.2) Binding Partners
Khanna, Rajesh ; Zougman, Alexandre ; Stanley, Elise F. ;
BMB Reports , volume 40, issue 3, 2007, Pages 302~314
DOI : 10.5483/BMBRep.2007.40.3.302
N type calcium channels (CaV2.2) play a key role in the gating of transmitter release at presynaptic nerve terminals. These channels are generally regarded as parts of a multimolecular complex that can modulate their open probability and ensure their location near the vesicle docking and fusion sites. However, the proteins that comprise this component remain poorly characterized. We have carried out the first open screen of presynaptic CaV2.2 complex members by an antibody-mediated capture of the channel from purified rat brain synaptosome lysate followed by mass spectroscopy. 589 unique peptides resulted in a high confidence match of 104 total proteins and 40 synaptosome proteome proteins. This screen identified several known CaV2.2 interacting proteins including syntaxin 1, VAMP, protein phosphatase 2A,
and spectrin and also a number of novel proteins, including clathrin, adaptin, dynamin, dynein, NSF and actin. The unexpected proteins were classified within a number of functional classes that include exocytosis, endocytosis, cytoplasmic matrix, modulators, chaperones, and cell-signaling molecules and this list was contrasted to previous reports that catalogue the synaptosome proteome. The failure to detect any postsynaptic density proteins suggests that the channel itself does not exhibit stable trans-synaptic attachments. Our results suggest that the channel is anchored to a cytoplasmic matrix related to the previously described particle web.
Nucleotide Sequence, Structural Investigation and Homology Modeling Studies of a Ca
-independent α-amylase with Acidic pH-profile
Sajedi, Reza Hassan ; Taghdir, Majid ; Naderi-Manesh, Hossein ; Khajeh, Khosro ; Ranjbar, Bijan ;
BMB Reports , volume 40, issue 3, 2007, Pages 315~324
DOI : 10.5483/BMBRep.2007.40.3.315
-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of
and EDTA. Therefore, KRA acts as a
-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of
and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis
-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the
-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.
A Novel MAP Kinase Gene in Cotton (Gossypium hirsutum L.), GhMAPK, is Involved in Response to Diverse Environmental Stresses
Wang, Meimei ; Zhang, Ying ; Wang, Jian ; Wu, Xiaoliang ; Guo, Xingqi ;
BMB Reports , volume 40, issue 3, 2007, Pages 325~332
DOI : 10.5483/BMBRep.2007.40.3.325
The mitogen-activated protein kinase (MAPK) cascade is one of the major and evolutionally conserved signaling pathways and plays pivotal role in the regulation of stress and developmental signals in plants. Here, a novel gene, termed Gossypium hirsutum MAPK (GhMAPK), was isolated from cotton. The full-length cDNA of GhMAPK encodes for a 372 amino acid protein that contains all 11 of the MAPK conserved subdomains and the phosphorylationactivation motif, TEY. Amino acid sequence alignment revealed that GhMAPK shared high identity with group-C MAPK in plants and showed 83~89% similarities with MAPKs from Arabidopsis, apricot, pea, petunia, and tobacco. Southern blot analysis indicated that the GhMAPK belonged to a multygene family in cotton. Two introns were found within the region of genomic sequence. Northern blot analysis revealed that the transcripts of GhMAPK accumulated markedly when the cotton seedlings were subjected to various abiotic stimuli such as wounding, cold (4
), or salinity stress; Furthermore, GhMAPK was upregulated by the exogenous signaling molecules, such as salicylic acid (SA) and hydrogen peroxide (
), as well as pathogen attacks. These results indicate that the GhMAPK, which has a high degree of identity with group-C plant MAPKs, may also play an important role in response to environmental stresses.
Cloning and Expression of Cyclodextrin Glycosyltransferase Gene from Paenibacillus sp. T16 Isolated from Hot Spring Soil in Northern Thailand
Charoensakdi, Ratiya ; Murakami, Shuichiro ; Aoki, Kenji ; Rimphanitchayakit, Vichien ; Limpaseni, Tipaporn ;
BMB Reports , volume 40, issue 3, 2007, Pages 333~340
DOI : 10.5483/BMBRep.2007.40.3.333
Gene encoding cyclodextrin glycosyltransferase (CGTase), from thermotolerant Paenibacillus sp. T16 isolated from hot spring area in northern Thailand, was cloned and expressed in E. coli (JM109). The nucleotide sequences of both wild type and transformed CGTases consisted of 2139 bp open reading frame, 713 deduced amino acids residues with difference of 4 amino acid residues. The recombinant cells required 24 h culture time and a neutral pH for culture medium to produce compatible amount of CGTase compared to 72 h culture time and pH 10 for wild type. The recombinant and wild-type CGTases were purified by starch adsorption and phenyl sepharose column chromatography and characterized in parallel. Both enzymes showed molecular weight of 77 kDa and similar optimum pHs and temperatures with recombinant enzyme showing broader range. There were some significant difference in pH, temperature stability and kinetic parameters. The presence of high starch concentration resulted in higher thermostability in recombinant enzyme than the wild type. The recombinant enzyme was more stable at higher temperature and lower pH, with lower
for coupling reaction using cellobiose and cyclodextrins as substrates.
STP-C, an Oncoprotein of Herpesvirus saimiri Augments the Activation of NF-κB through Ubiquitination of TRAF6
Chung, Young-Hwa ; Jhun, Byung-Hak ; Ryu, Su-Chak ; Kim, Heui-Soo ; Kim, Cheol-Min ; Kim, Bong-Seok ; Kim, Young-Ok ; Lee, Sang-Jun ;
BMB Reports , volume 40, issue 3, 2007, Pages 341~348
DOI : 10.5483/BMBRep.2007.40.3.341
Herpesvirus saimiri (HVS), a member of the
-herpesvirus family, encodes an oncoprotein called Saimiri Transforming Protein (STP) which is required for lymphoma induction in non-human primates. Previous study has shown that STP-C, an oncoprotein of HVS, activates NF-
B signaling pathway. However, the detailed mechanism of STP-Cmediated NF-
B activation has not been reported yet. We first report that STP-C interacts with TRAF6 protein in vivo and in vitro and further investigation shows that
residue of STP-C is critical for binding to TRAF6. Introduction of ubiquitin together with STP-C augments NF-
B activity compared to that of STP-C expression alone. STP-C expression further induces ubiquitination of endogenous TRAF6. In addition, either a deubiquitination enzyme, CYLD or a dominant negative E2-conjugation enzyme reduced NF-
B activity in spite of the presence of STP-C, supporting that the interaction between STP-C and TRAF6 induces ubiquitination of TRAF6. NF-
B activation by STP-C through the ubiquitinated TRAF6 causes the increased production of IL-8, an inflammatory chemokine and the enhanced expression of costimulatory molecule ICAM, which might ultimately contribute cellular transformation by the exposure of HVS-infected cells with inflammatory microenvironment and chronic activation.
Molecular Analysis and Expression Patterns of the 14-3-3 Gene Family from Oryza Sativa
Yao, Yuan ; Du, Ying ; Jiang, Lin ; Liu, Jin-Yuan ;
BMB Reports , volume 40, issue 3, 2007, Pages 349~357
DOI : 10.5483/BMBRep.2007.40.3.349
The ubiquitous family of 14-3-3 proteins functions as regulators in a variety of physiological processes. Eight rice 14-3-3 genes, designated OsGF14a through h, were identified from an exhaustive search of the genome database. Comparisons of deduced amino acid sequences reveal a high degree of identity among members of the OsGF14 family and reported Arabidopsis 14-3-3 proteins. A phylogenetic study indicates that OsGF14s contain both
forms, which is also confirmed by a structural analysis of OsGF14 genes. Furthermore, transcripts of OsGF14b, OsGF14c, OsGF14d, OsGF14e, OsGF14f and OsGF14g were detected in rice tissues. Their different expression patterns, the different effects of environmental stresses and plant hormones on their transcription levels, and the different complementary phenotypes in yeast 14-3-3 mutants not only indicates that OsGF14s are responsive to various stress conditions and regulated by multiple signaling pathways, but also suggests that functional similarity and diversity coexist among the members of OsGF14 family.
A Novel Mannose-binding Tuber Lectin from Typhonium divaricatum (L.) Decne (family Araceae) with Antiviral Activity Against HSV-II and Anti-proliferative Effect on Human Cancer Cell Lines
Luo, Yongting ; Xu, Xiaochao ; Liu, Jiwei ; Li, Jian ; Sun, Yisheng ; Liu, Zhen ; Liu, Jinzhi ; Damme, Els Van ; Balzarini, Jan ; Bao, Jinku ;
BMB Reports , volume 40, issue 3, 2007, Pages 358~367
DOI : 10.5483/BMBRep.2007.40.3.358
A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95
g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7
6.8), Bre-04 (41.5
4.8), and Lu-04 (11.4
0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with
of 5.176 mg/ml and 3.054
g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.
Ghrelin is Present in Teeth
Aydin, Suleyman ; Ozercan, I brahim Hanefi ; Geckil, Hikmet ; Dagli, Ferda ; Aydin, Suna ; Kumru, Sinem ; Kilic, Nermin ; Sahin, I brahim ; Ozercan, Mehmet Resat ;
BMB Reports , volume 40, issue 3, 2007, Pages 368~372
DOI : 10.5483/BMBRep.2007.40.3.368
Ghrelin belongs to the family of a gut-brain hormone that promotes food intake and controls energy balance. Recently, it has also been shown to regulate bone formation directly. Dental tissue shares several functional, developmental and anatomical similarities with bone, and in the present study we have investigated the presence of ghrelin in 44 human teeth using immunocytochemistry and radioimmunoassay. Both methods showed that the hormone is present in canines and molars, mainly in the odontoblasts but also in the pulp. Ghrelin could potentially play interesting physiological roles in teeth.
Systematic Studies of 12S Seed Storage Protein Accumulation and Degradation Patterns during Arabidopsis Seed Maturation and Early Seedling Germination Stages
Li, Qing ; Wang, Bai-Chen ; Xu, Yu ; Zhu, Yu-Xian ;
BMB Reports , volume 40, issue 3, 2007, Pages 373~381
DOI : 10.5483/BMBRep.2007.40.3.373
Seed storage proteins (SSPs) are important for seed germination and early seedling growth. We studied the accumulation and degradation profiles of four major Arabidopsis 12S SSPs using a 2-DE scheme combined with mass spectrometric methods. On the 2-DE map of 23 dpa (days post anthesis) siliques, 48 protein spots were identified as putative full-length or partial
subunits. Only 9 of them were found in 12 dpa siliques with none in younger than 8 dpa siliques, indicating that the accumulation of 12S SSPs started after the completion of cell elongation processes both in siliques and in developing seeds. The length and strength of transcription activity for each gene determined the final contents of respective SSP. At the beginning of imbibition, 68 SSP spots were identified while only 2 spots were found at the end of the 4 d germination period, with
, subunits degraded more rapidly than the
subunits. The CRC
subunit was found to degrade from its C-terminus with conserved sequence motifs. Our data provide an important basis for understanding the nutritional value of developing plant seeds and may serve as a useful platform for other species.
A 43 kD Protein Isolated from the Herb Cajanus indicus L Attenuates Sodium Fluoride-induced Hepatic and Renal Disorders in Vivo
Manna, Prasenjit ; Sinha, Mahua ; Sil, Parames C. ;
BMB Reports , volume 40, issue 3, 2007, Pages 382~395
DOI : 10.5483/BMBRep.2007.40.3.382
The herb, Cajanus indicus L, is well known for its hepatoprotective action. A 43 kD protein has been isolated, purified and partially sequenced from the leaves of this herb. A number of in vivo and in vitro studies carried out in our laboratory suggest that this protein might be a major component responsible for the hepatoprotective action of the herb. Our successive studies have been designed to evaluate the potential efficacy of this protein in protecting the hepatic as well as renal tissues from the sodium fluoride (NaF) induced oxidative stress. The experimental groups of mice were exposed to NaF at a dose of 600 ppm through drinking water for one week. This exposure significantly altered the activities of the antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR) and the cellular metabolites such as reduced glutathione (GSH), oxidized glutathione (GSSG), total thiols, lipid peroxidation end products in liver and kidney compared to the normal mice. Intraperitoneal administration of the protein at a dose of 2 mg/kg body weight for seven days followed by NaF treatment (600 ppm for next seven days) normalized the activities of the hepato-renal antioxidant enzymes, the level of cellular metabolites and lipid peroxidation end products. Post treatment with the protein for four days showed that it could help recovering the damages after NaF administration. Time-course study suggests that the protein could stimulate the recovery of both the organs faster than natural process. Effects of a known antioxidant, vitamin E, and a non-relevant protein, bovine serum albumin (BSA) have been included in the study to validate the experimental data. Combining all, result suggests that NaF could induce severe oxidative stress both in the liver and kidney tissues in mice and the protein possessed the ability to attenuate that hepato-renal toxic effect of NaF probably via its antioxidant activity.
Expression of Bacillus subtilis proBA Genes and Reduction of Feedback Inhibition of Proline Synthesis Increases Proline Production and Confers Osmotolerance in Transgenic Arabidopsis
Chen, Mingqing ; Wei, Hongbo ; Cao, JunWei ; Liu, Ruijie ; Wang, Youliang ; Zheng, Congyi ;
BMB Reports , volume 40, issue 3, 2007, Pages 396~403
DOI : 10.5483/BMBRep.2007.40.3.396
Proline accumulation has been shown to correlate with tolerance to drought and salt stresses in plants. We attempt to introduce the wild-type, mutant, and fusion proBA genes derived from Bacillus subtilis into Arabidopsis thaliana under the control of a strong promoter cauliflower mosaic virus 35S (CaMV35S). The transgenic plants produced higher level of free proline than control and the overproduction of proline resulted in the increased tolerance to osmotic stress in transgenic plants. Besides, the mutation in proBA genes, which were proved to lead
-glutamyl kinase (
-GK) reduces sensitivity to the end-product inhibition and the fusion of proB and proA also result in increasing proline production and confer osmotolerance in transgenic lines.
Induction of RNA-mediated Resistance to Papaya Ringspot Virus Type W
Krubphachaya, Pongrit ; Juricek, Mila ; Kertbundit, Sunee ;
BMB Reports , volume 40, issue 3, 2007, Pages 404~411
DOI : 10.5483/BMBRep.2007.40.3.404
Transformation of cantaloupes with the coat protein (cp) gene of papaya ringspot virus type W (PRSV-W), Thai isolate, was used to introduce virus resistance. Binary vectors containing either the full length coat protein coding region under control of the 35S CaMV promoter(pSA1175), or the inverted-repeat of a coat protein coding region (pSA1304), were constructed and used for Agrobacteriummediated transformation of cotyledonary explants of the cantaloupe cultivar Sun Lady. Four independent transgenic lines were obtained using pSA1304 and one using pSA1175. Integration of the PRSV-W cp gene into the genome of these transgenic lines was verified by PCR amplification, GUS assays and Southern blot hybridization. In vitro inoculation of these lines with PRSV-W revealed that whereas the line containing pSA1175 remained sensitive, the four lines containing pSA1304 were resistant. The presence of small RNA species, presumably siRNA, corresponding to regions of the viral cp gene in transgenic lines resistant to PRSV-W supports the involvement of post-transcriptional gene silencing in the establishment of resistance.
Purification and Physicochemical Characterization of a Recombinant Phospholipid Hydroperoxide Glutathione Peroxidase from Oryza sativa
Wang, Zebin ; Wang, Feng ; Duan, Rui ; Liu, Jin-Yuan ;
BMB Reports , volume 40, issue 3, 2007, Pages 412~418
DOI : 10.5483/BMBRep.2007.40.3.412
Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is an unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes. In the present work, the entire encoding region for Oryza sativa PHGPx was expressed in Escherichia coli M15, and the purified fusion protein showed a single band with 21.0 kD and pI = 8.5 on SDS- and IFE-PAGE, respectively. Judging from CD and fluorescence spectroscopy, this protein is considered to have a well-ordered structure with 12.2%
-turn, and 38.5% random coil. The optimum pH and temperature of the enzyme activity were pH 9.3 and 27
. The enzyme exhibited the highest affinity and catalytical efficiency to phospholipid hydroperoxide employing GSH or Trx as electron donor. Moreover, the protein displayed higher GSH-dependent activity towards t-Butyl-OOH and
. These results show that OsPHGPx is an enzyme with broad specificity for hydroperoxide substrates and yielded significant insight into the physicochemical properties and the dynamics of OsPHGPx.
Directed Evolution of Beta-galactosidase from Escherichia coli into Beta-glucuronidase
Xiong, Ai-Sheng ; Peng, Ri-He ; Zhuang, Jing ; Liu, Jin-Ge ; Xu, Fang ; Cai, Bin ; Guo, Zhao-Kui ; Qiao, Yu-Shan ; Chen, Jian-Min ; Zhang, Zhen ; Yao, Quan-Hong ;
BMB Reports , volume 40, issue 3, 2007, Pages 419~425
DOI : 10.5483/BMBRep.2007.40.3.419
In vitro directed evolution through DNA shuffling is a powerful molecular tool for creation of new biological phenotypes. E. coli
-glucuronidase are widely used, and their biological function, catalytic mechanism, and molecular structures are well characterized. We applied an in vitro directed evolution strategy through DNA shuffling and obtained five mutants named YG6764, YG6768, YG6769, YG6770 and YG6771 after two rounds of DNA shuffling and screening, which exhibited more
-glucuronidase activity than wild-type
-galactosidase. These variants had mutations at fourteen nucleic acid sites, resulting in changes in ten amino acids: S193N, T266A, Q267R, V411A, D448G, G466A, L527I, M543I, Q626R and Q951R. We expressed and purified those mutant proteins. Compared to the wild-type protein, five mutant proteins exhibited high
-glucuronidase activity. The comparison of molecular models of the mutated and wildtype enzymes revealed the relationship between protein function and structural modification.
Molecular Cloning of Hemoglobin Alpha-chain Gene from Pantholops hodgsonii, a Hypoxic Tolerance Species
Yingzhong, Yang ; Droma, Yunden ; Guoen, Jin ; Zhenzhong, Bai ; Lan, Ma ; Haixia, Yun ; Yue, Cao ; Kubo, Keishi ; Rili, Ge ;
BMB Reports , volume 40, issue 3, 2007, Pages 426~431
DOI : 10.5483/BMBRep.2007.40.3.426
To investigate the possible mechanisms of high-altitude native animals in adapting to high altitude, we cloned hemoglobin alpha-chain (alpha-chain Hb) gene from Pantholops hodgsonii, an animal species that indigenously lives at elevations of 3700-5500 m on the Qinghai-Tibetan plateau. Using reverse transcription polymerase chain reaction (RT-PCR) technique, the alpha-chain Hb gene was amplified from total RNA in the liver of the Pantholops hodgsonii. TA cloning technique was used and the PCR product was cloned into pGEM-T vector. The DNA sequence of the gene was highly homologous with sheep (99.1%), goat (98.6%), cattle (95.6%) and human (86.5%). The alpha-chain Hb gene encoded a 142-amino acid protein that could be identified with the homology of alpha-chain Hb protein in sheep (98%), goat (96%), cattle (91%) and human (87%). However, 18 alternations were detected when compared with the alpha-chain Hb gene in human, and 2 in sheep. Moreover, the alterations of a117 GluAsp and
132 AsnSer in important regions were noted in human and sheep, respectively. Phylogenetic analysis suggested that the structure of alpha-chain Hb was highly similar to that in sheep. This study provided essential information for elucidating the possible roles of hemoglobin in adapting to extremely high altitude in Pantholops hodgsonii.
DNA-dependent Protein Kinase Mediates V(D)J Recombination via RAG2 Phosphorylation
Hah, Young-Sool ; Lee, Jung-Hwa ; Kim, Deok-Ryong ;
BMB Reports , volume 40, issue 3, 2007, Pages 432~438
DOI : 10.5483/BMBRep.2007.40.3.432
V(D)J recombination, a site-specific gene rearrangement process occurring during the lymphocyte development, begins with DNA double strand breaks by two recombination activating gene products (RAG1/2) and finishes with the repair process by several proteins including DNA-dependent protein kinase (DNA-PK). In this report, we found that RAG2 was specifically phosphorylated by DNA-PK at the
serine residue, and this phosphorylated RAG2 affected the V(D)J recombination activity in cells in the GFP expression-based assay. While the V(D)J recombination activity between wild-type RAG2 and mutant S365A RAG2 in the assay using a signal joint substrate was undistinguishable in DNA-PK deficient cells (M059J), the activity with wild-type RAG2 was largely increased in DNA-PK proficient cells (M059K) in comparison with mutant RAG2, suggesting that RAG2 phosphorylation by DNA-PK plays a crucial role in the signal joint formation during V(D)J recombination.
Saxatilin Suppresses Tumor-induced Angiogenesis by Regulating VEGF Expression in NCI-H460 Human Lung Cancer Cells
Jang, Yoon-Jung ; Kim, Dong-Seok ; Jeon, Ok-Hee ; Kim, Doo-Sik ;
BMB Reports , volume 40, issue 3, 2007, Pages 439~443
DOI : 10.5483/BMBRep.2007.40.3.439
Tumor growth and metastasis are dependent on angiogenesis, and endothelial cell invasion and migration are apparent means of regulating tumor progression. We report here that saxatilin, a snake venom-derived disintegrin, suppresses the angiogenesis-inducing properties of NCI-H460 human lung cancer cells. Culture supernatants of NCI-H460 cells are able to induce human umbilical vascular endothelial cell (HUVEC) invasion and tube formation. However, treatment of the cancer cells with saxatilin resulted in reduced angiogenic activity of the culture supernatant. This suppressed angiogenic property was found to be associated with the level of vascular endothelial growth factor (VEGF) in the culture supernatant. Further experimental evidence indicated that saxatilin inhibits VEGF production in NCI-H460 cells by affecting hypoxia induced factor-1
) expression via the Akt pathway.
A Simple and Rapid Gene Amplification from Arabidopsis Leaves Using AnyDirect System
Yang, Young-Geun ; Kim, Jong-Yeol ; Soh, Moon-Soo ; Kim, Doo-Sik ;
BMB Reports , volume 40, issue 3, 2007, Pages 444~447
DOI : 10.5483/BMBRep.2007.40.3.444
Polymerase chain reaction (PCR) is a powerful technique in molecular biology and is widely used in various fields. By amplifying DNA fragments, PCR has facilitated gene cloning procedures, as well as molecular genotyping. However, the extraction of DNA from samples often acts as a limiting step of these reactions. In particular, the extraction of PCR-compatible genomic DNA from higher plants requires complicated processes and tedious work because plant cells have rigid cell walls and contain various endogenous PCR inhibitors, including polyphenolic compounds. We recently developed a novel solution, referred to as AnyDirect, which can amplify target DNA fragments directly from whole blood without the need for DNA extraction. Here, we developed a simple lysis system that could produce an appropriate template for direct PCR with AnyDirect PCR buffer, making possible the direct amplification of DNA fragments from plant leaves. Thus, our experimental procedure provides a simple, convenient, non-hazardous, inexpensive, and rapid process for the amplification of DNA from plant tissue.
Contributions of CYP2C9/CYP2C19 Genotypes and Drug Interaction to the Phenytoin Treatment in the Korean Epileptic Patients in the Clinical Setting
Lee, Soo-Youn ; Lee, Seung-Tae ; Kim, Jong-Won ;
BMB Reports , volume 40, issue 3, 2007, Pages 448~452
DOI : 10.5483/BMBRep.2007.40.3.448
We examined the contribution of CYP2C9 and CYP2C19 genotypes and drug interactions to the phenytoin metabolism among 97 Korean epileptic patients to determine if pharmacogenetic testing could be utilized in routine clinical practice. The CYP2C9 polymorphism is a wellknown major genetic factor responsible for phenytoin metabolism. The CYP219 polymorphism, with a high incidence of variant alleles, has a minor influence on phenytoin treated Koran patients. Using a multiple regression model for evaluation of the CYP2C9 and CYP2C19 genotypes, together with other non-genetic variables, we explained 39.6% of the variance in serum phenytoin levels. Incorporation of genotyping for CYP2C9 and CYP2C19 into a clinical practice may be of some help in the determination of phenytoin dosage. However, because concurrent drug treatment is common in patients taking phenytoin and many environmental factors are likely to play a role in drug metabolism, these factors may overwhelm the relevance of CYP polymorphisms in the clinical setting. Further investigations with an approach to dose assessment that includes comprehensive interpretation of both pharmacogenetic and pharmacokinetic data along with understanding of the mechanism of drug interactions in dosage adjustment is warranted.