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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Korean Society for Biochemistry and Molecular Biology
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Volume & Issues
Volume 41, Issue 12 - Dec 2008
Volume 41, Issue 11 - Nov 2008
Volume 41, Issue 10 - Oct 2008
Volume 41, Issue 9 - Sep 2008
Volume 41, Issue 8 - Aug 2008
Volume 41, Issue 7 - Jul 2008
Volume 41, Issue 6 - Jun 2008
Volume 41, Issue 5 - May 2008
Volume 41, Issue 4 - Apr 2008
Volume 41, Issue 3 - Mar 2008
Volume 41, Issue 2 - Feb 2008
Volume 41, Issue 1 - Jan 2008
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Blood-neural barrier: its diversity and coordinated cell-to-cell communication
Choi, Yoon-Kyung ; Kim, Kyu-Won ;
BMB Reports , volume 41, issue 5, 2008, Pages 345~352
DOI : 10.5483/BMBRep.2008.41.5.345
The cerebral microvessels possess barrier characteristics which are tightly sealed excluding many toxic substances and protecting neural tissues. The specialized blood-neural barriers as well as the cerebral microvascular barrier are recognized in the retina, inner ear, spinal cord, and cerebrospinal fluid. Microvascular endothelial cells in the brain closely interact with other components such as astrocytes, pericytes, perivascular microglia and neurons to form functional `neurovascular unit`. Communication between endothelial cells and other surrounding cells enhances the barrier functions, consequently resulting in maintenance and elaboration of proper brain homeostasis. Furthermore, the disruption of the neurovascular unit is closely involved in cerebrovascular disorders. In this review, we focus on the location and function of these various blood-neural barriers, and the importance of the cell-to-cell communication for development and maintenance of the barrier integrity at the neurovascular unit. We also demonstrate the close relation between the alteration of the blood-neural barriers and cerebrovascular disorders.
Application of hybrid LRR technique to protein crystallization
Jin, Mi-Sun ; Lee, Jie-Oh ;
BMB Reports , volume 41, issue 5, 2008, Pages 353~357
DOI : 10.5483/BMBRep.2008.41.5.353
LRR family proteins play important roles in a variety of physiological processes. To facilitate their production and crystallization, we have invented a novel method termed "Hybrid LRR Technique". Using this technique, the first crystal structures of three TLR family proteins could be determined. In this review, design principles and application of the technique to protein crystallization will be summarized. For crystallization of TLRs, hagfish VLR receptors were chosen as the fusion partners and the TLR and the VLR fragments were fused at the conserved LxxLxLxxN motif to minimize local structural incompatibility. TLR-VLR hybridization did not disturb structures and functions of the target TLR proteins. The Hybrid LRR Technique is a general technique that can be applied to structural studies of other LRR proteins. It may also have broader application in biochemical and medical application of LRR proteins by modifying them without compromising their structural integrity.
Multiple shRNA expressing vector enhances efficiency of gene silencing
Song, Jun ; Giang, An ; Lu, Yingchun ; Pang, Shen ; Chiu, Robert ;
BMB Reports , volume 41, issue 5, 2008, Pages 358~362
DOI : 10.5483/BMBRep.2008.41.5.358
RNA interference (RNAi) is the process of sequence-specific gene silencing. However, RNAi efficiency still needs to be improved for effective inhibition of target genes. We have developed an effective strategy to express multiple shRNAs (small hairpin RNA) simultaneously using multiple RNA Polymerase III (Pol III) promoters in a single vector. Our data demonstrate that multiple shRNAs expressed from Pol III promoters have a synergistic effect in repressing the target gene. Silencing of endogenous cyclophilin A (CypA) or key HIV viral genes by multiple shRNAs results in significant inhibition of the target gene.
Overexpression of a delayed early gene hlg1 of temperate mycobacteriophage L1 is lethal to both M. smegmatis and E. coli
Chattoraj, Partho ; Ganguly, Tridib ; Nandy, Ranjan Kumar ; Sau, Subrata ;
BMB Reports , volume 41, issue 5, 2008, Pages 363~368
DOI : 10.5483/BMBRep.2008.41.5.363
Two genes of temperate mycobacteriophage L5, namely, gp63 and gp64, were hypothesized to be toxic to M. smegmatis. An identical L5 gp64 ortholog (designated hlg1) was cloned from homoimmune mycobacteriophage L1 and characterized at length here. As expected, hlg1 affected the growth of M. smegmatis when overexpressed from a resident plasmid. HLG1 (the protein encoded by hlg1) in fact caused growth retardation of M. smegmatis and the region encompassing its 57-114 C-terminal amino acid residues was found indispensable for its growthretardation activity. Both nucleic acid and protein biosynthesis were severely impaired in M. smegmatis expressing HLG1. Interestingly, HLG1 also affected E. coli almost similarly. This putative delayed early lipoprotein did not participate in the lytic growth of L1.
Apoptosis-inducing effect and structural basis of Polygonatum cyrtonema lectin and chemical modification properties on its mannose-binding sites
Liu, Bo ; Xu, Xiao-Chao ; Cheng, Yan ; Huang, Jian ; Liu, Yan-Hong ; Liu, Zhen ; Min, Ming-Wei ; Bian, He-Jiao ; Che, Jing ; Bao, Jin-Ku ;
BMB Reports , volume 41, issue 5, 2008, Pages 369~375
DOI : 10.5483/BMBRep.2008.41.5.369
Polygonatum cyrtonema Lectin (PCL), which is classified as a monocot mannose-binding lectin, has received great regards for its uniquely biological activities and potentially medical applications in cancer cells. This paper was initially aimed to study apoptosis of PCL on Hela cells. Thus, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method was carried out. Through observation of cell morphologic changes and Lactate dehydrogenase (LDH) activity-based cytotoxicity assays, PCL induced HeLa cell apoptosis in a dose-dependent manner. To further gain structural basis, multiple alignments, homology modeling and docking experiments were performed to analyze the correlation between its biological activities and mannose-binding sites. Eventually, considering docking data, chemical modification properties on the three mannose-binding sites were analyzed by a series of biological experiments (e.g., hemagglutinating and mitogenic activity assays, fluorescence and Circular Dichrosim (CD) spectroscopy) to profoundly identify the role of some key amino acids in the structure-function relationship of PCL.
RNA silencing-mediated resistance is related to biotic / abiotic stresses and cellular RdRp expression in transgenic tobacco plants
Wu, Xiao-Liang ; Hou, Wen-Cui ; Wang, Mei-Mei ; Zhu, Xiao-Ping ; Li, Fang ; Zhang, Jie-Dao ; Li, Xin-Zheng ; Guo, Xing-Qi ;
BMB Reports , volume 41, issue 5, 2008, Pages 376~381
DOI : 10.5483/BMBRep.2008.41.5.376
The discovery of RNA silencing inhibition by virus encoded suppressors or low temperature leads to concerns about the stability of transgenic resistance. RNA-dependent RNA polymerase (RdRp) has been previously characterized to be essential for transgene-mediated RNA silencing. Here we showed that low temperature led to the inhibition of RNA silencing, the loss of viral resistance and the reduced expression of host RdRp homolog (NtRdRP1) in transgenic T4 progeny with untranslatable potato virus Y coat protein (PVY-CP) gene. Moreover, RNA silencing and the associated resistance were differently inhibited by potato virus X (PVX) and tobacco mosaic virus (TMV) infections. The increased expression of NtRdRP1 in both PVX and TMV infected plants indicated its general role in response to viral pathogens. Collectively, we propose that biotic and abiotic stress factors affect RNA silencing-mediated resistance in transgenic tobacco plants and that their effects target different steps of RNA silencing.
Heat stress protection in Aspen sp1 transgenic Arabidopsis thaliana
Zhu, Bo ; Xiong, Ai-Sheng ; Peng, Ri-He ; Xu, Jing ; Zhou, Jun ; Xu, Jin-Tao ; Jin, Xiao-Fen ; Zhang, Yang ; Hou, Xi-Lin ; Yao, Quan-Hong ;
BMB Reports , volume 41, issue 5, 2008, Pages 382~387
DOI : 10.5483/BMBRep.2008.41.5.382
It is known that the stable protein 1 (SP1) detected in aspen plants remains soluble upon boiling and that sp1 expression in transgenic aspen is resistant to salt stress. Presently, we analyzed the effect of expression of SP1 in Arabidopsis thaliana plants and their response to high temperature stress. After
for 16 h, relative to wild type plants, sp1 transgenic plants exhibited stronger growth and were better in several physiological properties including chlorophyII, chlorophyII fluorescence, water content, proline content, and malondialdehyde content. These preliminarily results suggest that the over-expression of SP1 may notably enhance heat-tolerant level of transgenic A. thaliana plants.
A heat shock cognate 70 gene in the endoparasitoid, Pteromalus puparum, and its expression in relation to thermal stress
Wang, Huan ; Dong, Sheng-Zhang ; Li, Kai ; Hu, Cui ; Ye, Gong-Yin ;
BMB Reports , volume 41, issue 5, 2008, Pages 388~393
DOI : 10.5483/BMBRep.2008.41.5.388
The Pphsc70 (heat shock cognate 70) gene was isolated from the endoparasitoid Pteromalus puparum and then characterized. The full-length cDNA was 2204 base pair (bp) and contained a single 1968 bp ORF that encoded a polypeptide of 656 amino acids with a predicted molecular mass of 71.28 kDa. Phylogenetic analysis based on Hsc70 amino acid sequences from fifteen insect species agreed with the present phylogeny. In addition, genomic DNA confirmed the presence of three introns located at the coding region as well as the 5`UTR. A significant elevation of Pphsc70 expression was observed following heat treatment, however, continued exposure to heat shock or recovery caused the expression of induced mRNA to gradually decline to levels that were significantly lower than those of control pupae (P < 0.05). In addition, a significant increase was observed in the emergence rate of pupae that were preheated at
and then exposed to
for 1 h when compared with the pupae that were not preheated, but instead directly exposed to
. Taken together, these results revealed that exposure to gradually increasing temperatures can enhance an insects thermo-tolerance.
Intrinsic bent DNA colocalizes with the sequence involved in the Nd-s
mutation in the Bombyx mori fibroin light chain gene
Barbosa, Joice Felipes ; Bravo, Juliana Pereira ; Takeda, Karen Izumi ; Zanatta, Daniela Bertolini ; Silva, Jose Luis Da Conceicao ; Balani, Valerio Americo ; Fiorini, Adriana ; Fernandez, Maria Aparecida ;
BMB Reports , volume 41, issue 5, 2008, Pages 394~399
DOI : 10.5483/BMBRep.2008.41.5.394
Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd-
mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-
mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene.
Bombyx mori protein disulfide isomerase enhances the production of nuecin, an antibacterial protein
Goo, Tae-Won ; Yun, Eun-Young ; Kim, Sung-Wan ; Choi, Kwang-Ho ; Kang, Seok-Woo ; Kwon, Ki-Sang ; Yu, Kweon ; Kwon, O-Yu ;
BMB Reports , volume 41, issue 5, 2008, Pages 400~403
DOI : 10.5483/BMBRep.2008.41.5.400
The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpressions of foreign proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins.
Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli
Ihm, Hyo-Jin ; Yang, Seung-Ju ; Huh, Jae-Wan ; Choi, Soo-Young ; Cho, Sung-Woo ;
BMB Reports , volume 41, issue 5, 2008, Pages 404~407
DOI : 10.5483/BMBRep.2008.41.5.404
A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2.
Human brain pyridoxal-5`-phosphate phosphatase (PLPP): protein transduction of PEP-1-PLPP into PC12 cells
Lee, Yeom-Pyo ; Kim, Dae-Won ; Lee, Min-Jung ; Jeong, Min-Seop ; Kim, So-Young ; Lee, Sun-Hwa ; Jang, Sang-Ho ; Park, Jin-Seu ; Kang, Tae-Cheon ; Won, Moo-Ho ; Cho, Sung-Woo ; Kwon, Oh-Shin ; Eum, Won-Sik ; Choi, Soo-Young ;
BMB Reports , volume 41, issue 5, 2008, Pages 408~413
DOI : 10.5483/BMBRep.2008.41.5.408
Pyridoxal-5`-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5`-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin
precursors; pyridoxine, pyridoxal kinase and pyridoxine-5`-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin