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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 39, Issue 6 - Nov 2006
Volume 39, Issue 5 - Sep 2006
Volume 39, Issue 4 - Jul 2006
Volume 39, Issue 3 - May 2006
Volume 39, Issue 2 - Mar 2006
Volume 39, Issue 1 - Jan 2006
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Differential Roles of Vascular Endothelial Growth Factor Receptor-1 and Receptor-2 in Angiogenesis
Shibuya, Masabumi ;
BMB Reports , volume 39, issue 5, 2006, Pages 469~478
DOI : 10.5483/BMBRep.2006.39.5.469
Vascular endothelial growth factor (VEGF)-A, a major regulator for angiogenesis, binds and activates two tyrosine kinase receptors, VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1). These receptors regulate physiological as well as pathological angiogenesis. VEGFR2 has strong tyrosine kinase activity, and transduces the major signals for angiogenesis. However, unlike other representative tyrosine kinase receptors which use the Ras pathway, VEGFR2 mostly uses the Phospholipase-
-Protein kinase-C pathway to activate MAP-kinase and DNA synthesis. VEGFR2 is a direct signal transducer for pathological angiogenesis including cancer and diabetic retinopathy, thus, VEGFR2 itself and the signaling appear to be critical targets for the suppression of these diseases. VEGFR1 plays dual role, a negative role in angiogenesis in the embryo most likely by trapping VEGF-A, and a positive role in adulthood in a tyrosine kinase-dependent manner. VEGFR1 is expressed not only in endothelial cells but also in macrophage-lineage cells, and promotes tumor growth, metastasis, and inflammation. Furthermore, a soluble form of VEGFR1 was found to be present at abnormally high levels in the serum of preeclampsia patients, and induces proteinurea and renal dysfunction. Therefore, VEGFR1 is also an important target in the treatment of human diseases. Recently, the VEGFR2-specific ligand VEGF-E (Orf-VEGF) was extensively characterized. Interestingly, the activation of VEGFR2 via VEGF-E in vivo results in a strong angiogenic response in mice with minor side effects such as inflammation compared with VEGF-A, suggesting VEGF-E to be a novel material for pro-angiogenic therapy.
Heme Oxygenase-1 as a Potential Therapeutic Target for Hepatoprotection
Farombi, Ebenezer Olatunde ; Surh, Young-Joon ;
BMB Reports , volume 39, issue 5, 2006, Pages 479~491
DOI : 10.5483/BMBRep.2006.39.5.479
Heme oxygenase (HO), the rate limiting enzyme in the breakdown of heme into carbon monoxide (CO), iron and bilirubin, has recently received overwhelming research attention. To date three mammalian HO isozymes have been identified, and the only inducible form is HO-1 while HO-2 and HO-3 are constitutively expressed. Advances in unveiling signal transduction network indicate that a battery of redox-sensitive transcription factors, such as activator protein-1 (AP-1), nuclear factor-kappa B (NF-
) and nuclear factor E2-related factor-2 (Nrf2), and their upstream kinases including mitogen-activated protein kinases play an important regulatory role in HO-1 gene induction. The products of the HO-catalyzed reaction, particularly CO and biliverdin/bilirubin have been shown to exert protective effects in several organs against oxidative and other noxious stimuli. In this context, it is interesting to note that induction of HO-1 expression contributes to protection against liver damage induced by several chemical compounds such as acetaminophen, carbon tetrachloride and heavy metals, suggesting HO-1 induction as an important cellular endeavor for hepatoprotection. The focus of this review is on the significance of targeted induction of HO-1 as a potential therapeutic strategy to protect against chemically-induced liver injury as well as hepatocarcinogenesis.
GTP Induces S-phase Cell-cycle Arrest and Inhibits DNA Synthesis in K562 Cells But Not in Normal Human Peripheral Lymphocytes
Moosavi, Mohammad Amin ; Yazdanparast, Razieh ; Lotfi, Abbas ;
BMB Reports , volume 39, issue 5, 2006, Pages 492~501
DOI : 10.5483/BMBRep.2006.39.5.492
Since differentiation therapy is one of the promising strategies for treatment of leukemia, universal efforts have been focused on finding new differentiating agents. In that respect, we used guanosine 5'-triphosphate (GTP) to study its effects on K562 cell line. GTP, at concentrations between 25-200
, inhibited proliferation (3-90%) and induced 5-78% increase in benzidine-positive cells after 6-days of treatments of K562 cells. Flow cytometric analyses of glycophorine A (GPA) showed that GTP can induce expression of this marker in more mature erythroid cells in a time- and dose-dependent manner. These effects of GTP were also accompanied with inhibition of DNA synthesis (measured by [
]-thymidine incorporation) and early S-phase cell cycle arrest by 96 h of exposure. In contrast, no detectable effects were observed when GTP administered to unstimulated human peripheral blood lymphocytes (PBL). However, GTP induced an increase in proliferation, DNA synthesis and viability of mitogen-stimulated PBL cells. In addition, growth inhibition and differentiating effects of GTP were also induced by its corresponding nucleotides GDP, GMP and guanosine (Guo). In heat-inactivated medium, where rapid degradation of GTP via extracellular nucleotidases is slow, the anti-proliferative and differentiating effects of all type of guanine nucleotides (except Guo) were significantly decreased. Moreover, adenosine, as an inhibitor of Guo transporter system, markedly reduced the GTP effects in K562 cells, suggesting that the extracellulr degradation of GTP or its final conversion to Guo may account for the mechanism of GTP effects. This view is further supported by the fact that GTP and Guo are both capable of impeding the effects of mycophenolic acid. In conclusion, our data will hopefully have important impact on pharmaceutical evaluation of guanine nucleotides for leukemia treatments.
Molecular Cloning, Characterization and Functional Analysis of a 2C-methyl-D-erythritol 2, 4-cyclodiphosphate Synthase Gene from Ginkgo biloba
Gao, Shi ; Lin, Juan ; Liu, Xuefen ; Deng, Zhongxiang ; Li, Yingjun ; Sun, Xiaofen ; Tang, Kexuan ;
BMB Reports , volume 39, issue 5, 2006, Pages 502~510
DOI : 10.5483/BMBRep.2006.39.5.502
2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase (MECPS, EC: 184.108.40.206) is the fifth enzyme of the non-mevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate (MEP) pathway for ginkgolide biosynthesis. The full-length mecps cDNA sequence (designated as Gbmecps) was cloned and characterized for the first time from gymnosperm plant species, Ginkgo biloba, using RACE (rapid amplification of cDNA ends) technique. The full-length cDNA of Gbmecps was 874 bp containing a 720 bp open reading frame (ORF) encoding a peptide of 239 amino acids with a calculated molecular mass of 26.03 kDa and an isoelectric point of 8.83. Comparative and bioinformatic analyses revealed that GbMECPS showed extensive homology with MECPSs from other species and contained conserved residues owned by the MECPS protein family. Phylogenetic analysis indicated that GbMECPS was more ancient than other plant MECPSs. Tissue expression pattern analysis indicated that GbMECPS expressed the highest in roots, followed by in leaves, and the lowest in seeds. The color complementation assay indicated that GbMECPS could accelerate the accumulation of
-carotene. The cloning, characterization and functional analysis of GbMECPS will be helpful to understand more about the role of MECPS involved in the ginkgolides biosynthesis at the molecular level.
Alanine Aminotransferase in Amphioxus: Presence, Localization and Up-regulation after Acute Lipopolysaccharide Exposure
Lun, Li-Min ; Zhang, Shi-Cui ; Liang, Yu-Jun ;
BMB Reports , volume 39, issue 5, 2006, Pages 511~515
DOI : 10.5483/BMBRep.2006.39.5.511
Alanine aminotransferase (AAT) is mainly synthesized in the liver, and its level in mammalian serum is elevated after acute phase induction. Here we demonstrated that sheep anti-human AAT antibody cross-reacted with amphioxus humoral fluids as well as human serum; and the concentration of AAT in the humoral fluids in amphioxus increased after the acute challenge with lipopolysaccharide, while the level of total proteins remains unchanged. These suggest the presence of the same acute phase response pattern in amphioxus, as observed in some mammalian species. Immunohistochemically, AAT was localized in the hepatic diverticulum, ovary and testis. It appears that the hepatic diverticulum in amphioxus is functionally homologous to the vertebrate liver in respect of AAT synthesis, supporting the hypothesis that the vertebrate liver evolved from the hepatic diverticulum of an amphioxus-like ancestor during early chordate evolution.
Determination of Saturated and Unsaturated Fatty Acids Amount in Leukocyte Membranes from Subjects Fed with Solid and Fluid Oils
Erman, Fazilet ; Aydin, Suleyman ; Demir, Yasar ; Akcay, Fatih ; Bakan, Ebubekir ;
BMB Reports , volume 39, issue 5, 2006, Pages 516~521
DOI : 10.5483/BMBRep.2006.39.5.516
Modifications in dietary fatty acid intake might lead to a modification in membrane phospholipid fatty acid composition. The purpose of this study was to investigate relationship between different type of oil consumption and leukocyte membrane phospholipid composition. This study was carried out in subjects utilizing butter (n = 15), margarine (n = 15), fluid oil (n = 15) and mixed types of oils (n = 15) in total 60 subjects. Leukocytes were separated from total blood by dextran sedimentation method. Membrane lipids and proteins were isolated following the cell disruption. Fatty acids of membrane phospholipids were isolated by hydrolysation with phospholipase B under ultrasonic dismembranator. Free fatty acids were identified with gas chromatography at chloroform phase. The results obtained were compared with data obtained by chromatograms of the standards. Results more prominent values of arachidic, dihomo-
-linolenic and palmitoleic acids were found in butter-or mixed oil-user groups; eicosadienoic, eicosamonoenoic, dihomo-
-linolenic and behenic acids in fluid oil; heptanoic, valeric, eicosadienoic and linolenic acids in margarine groups. The fatty acid composition of mixed oil was similar to butter, while other two oils were so different. From this study, it was concluded that the type of oil consumption might have an influence on phospholipid components of plasma membranes.
Molecular Phylogeny of Silk Producing Insects Based on Internal Transcribed Spacer DNA1
Mahendran, Botlagunta ; Ghosh, Sudip K. ; Kundu, Subhas C. ;
BMB Reports , volume 39, issue 5, 2006, Pages 522~529
DOI : 10.5483/BMBRep.2006.39.5.522
Silk moths are the best studied silk secreting insects and belong to the families Bombycidae and Saturniidae. The phylogenetic relationship between eleven silk producing insects was analyzed using the complete DNA sequence of the internal transcribed spacer DNA 1 locus. The PCR amplification and sequence analysis showed variation in length ranging from 138 bp (Antheraea polyphemus) to 911 bp (Hyalopora cecropia). Microsatellite sequences were found and was be used to distinguish Saturniidae and Bombycidae members. The nucleotide sequences were aligned manually and used for construction of phylogenetic trees based on Maximum parsimony and Maximum likelihood methods. The topology in both the approaches yielded a similar tree that supports the ancestral position of the Antheraea assama.
Conformational Study of Human Serum Albumin in Pre-denaturation Temperatures by Differential Scanning Calorimetry, Circular Dichroism and UV Spectroscopy
Rezaei-Tavirani, Mostafa ; Moghaddamnia, Seyed Hassan ; Ranjbar, Bijan ; Amani, Mojtaba ; Marashi, Sayed-Amir ;
BMB Reports , volume 39, issue 5, 2006, Pages 530~536
DOI : 10.5483/BMBRep.2006.39.5.530
Thermal conformational changes of human serum albumin (HSA) in phosphate buffer, 10 mM at pH = 7 are investigated using differential scanning calorimetric (DSC), circular dichroism (CD) and UV spectroscopic methods. The results indicate that temperature increment from
induces reversible conformational changes in the structure of HSA. Conformational change of HSA are shown to be a three-step process. Interestingly, melting temperature of the last domain is equal to the maximum value of fever in pathological conditions, i.e.
. These conformational alterations are accompanied by a mild alteration of secondary structures. Study of HSA-SDS (sodium dodecyl sulphate) interaction at
reveals that SDS affects the HSA structure at least in three steps: the first two steps result in more stabilization and compactness of HSA structure, while the last one induces the unfolding of HSA. Since HSA has a more affinity for SDS at
, It is suggested that the net negative charge of HSA is decreased in fever, which results in the decrease of HSA-associated cations and plasma osmolarity, and consequently, heat removal via the increase in urine volume.
Characterization and Expression Profile of CMTM3/CKLFSF3
Zhong, Ji ; Wang, Yu ; Qiu, Xiaoyan ; Mo, Xiaoning ; Liu, Yanan ; Li, Ting ; Song, Quansheng ; Ma, Dalong ; Han, Wenling ;
BMB Reports , volume 39, issue 5, 2006, Pages 537~545
DOI : 10.5483/BMBRep.2006.39.5.537
CMTM/CKLFSF is a novel family of proteins linking chemokines and TM4SF. In humans, these proteins are encoded by nine genes, CKLF and CMTM1-8/CKLFSF1-8. Here we report the characteristics and expression profile of CMTM3/CKLFSF3. Human CMTM3/CKLFSF3 has a high sequence identity among various species and similar characteristics as its mouse and rat homologues. Established by results both of RT-PCR and Quantitative Real-time PCR, the gene is highly transcribed in testis, leukocytes and spleen. For further verification, we generated a polyclonal antibody against human CMTM3/CKLFSF3 and found that the protein is highly expressed in the testis and some cells of PBMCs. Therefore, CMTM3/CKLFSF3 is an evolutionarily conserved gene that may have important roles in the male reproductive system and immune system. Further studies are necessary to validate its functions in the two systems.
Purification and Partial Characterization of an Acidic Polysaccharide with Complement Fixing Ability from the Stems of Avicennia Marina
Fang, Xubo ; Jiang, Bo ; Wang, Xiaolan ;
BMB Reports , volume 39, issue 5, 2006, Pages 546~555
DOI : 10.5483/BMBRep.2006.39.5.546
An acidic polysaccharide fraction that had high anti-complementary activity was isolated from the stems of Grey Mangrove in 0.15% yield. The final fractions was designated HAM-3-IIb-II. The polysaccharide fraction appeared to be homogenous by high performance size exclusion chromatography with an estimated molecular weight of 105 kDa. The isolated polysaccharide is more effective than polysaccharide K (PSK) in its anti-complementary activity at 58
of PSK and 23
of HAM-3-IIb-II that inhibit 50% of complement activity in the complement fixation assay. Structural studies indicated that HAM-3-IIb-II was rich in galacturonic acid along with arabinose, galactose and rhamnose, characterizing a pectin-type polysaccharide, which was also confirmed by FT-IR spectrum. The presence of rich neutral sugar side chains of arabinogalactans may have contributed to the expression of high activity. Traditionally, this mangrove plant is used for medicinal purposes and it appears to have some scientific applications.
Differential Efflux of Mitochondrial Endonuclease G by hNoxa and tBid
Seo, Young-Woo ; Park, Sun-Young ; Yun, Cheol-Won ; Kim, Tae-Hyoung ;
BMB Reports , volume 39, issue 5, 2006, Pages 556~559
DOI : 10.5483/BMBRep.2006.39.5.556
The Bcl-2 family of proteins regulates mitochondrial functions during cell death by modulating the efflux of death-promoting proteins such as cytochrome c and endonuclease G. Upon the binding of death ligands to their receptors, caspase-8 cleaves Bid, a BH3-only protein, into tBid that causes the mitochondrial damages resulting in the release of cytochrome c and endonuclease G. Also, another BH3-only protein, hNoxa, has been shown to induce the efflux of cytochrome c from the mitochondria. Whether the efflux proteins from the mitochondria in response to tBid or hNoxa are the same or different, however, has not been addressed. We have demonstrated that endonuclease G activities are not detectable among the proteins released from isolated mitochondria by hNoxa but are detectable in that by tBid. These results suggest that the efflux of proteins from the mitochondria are differentially modulated by tBid and hNoxa.
Molecular Characterization of the Recombinant A-chain of a Type II Ribosome-Inactivating Protein (RIP) from Viscum album coloratum and Structural Basis on its Ribosome-Inactivating Activity and the Sugar-binding Properties of the B-chain
Ye, Wenhui ; Nanga, Ravi Prakash Reddy ; Kang, Cong Bao ; Song, Joo-Hye ; Song, Seong-Kyu ; Yoon, Ho-Sup ;
BMB Reports , volume 39, issue 5, 2006, Pages 560~570
DOI : 10.5483/BMBRep.2006.39.5.560
Mistletoe (Viscum album) lectins, which are classified as a type II ribosome-inactivating protein (RIP) due to their unique biological function and the potential medical and therapeutic application in cancer cells, receive a rising attention. The heterodimeric glycoproteins contain the A-chain with catalytic activity and the B-chain with sugar binding properties. In recent years, studies involving the lectins from the white berry European mistletoe (Viscum album) and the yellow berry Korean mistletoe (Viscum album coloratum) have been described. However, the detailed mechanism in exerting unique cytotoxic effect on cancer cells still remains unclear. Here, we aim to understand and define the molecular basis and biological effects of the type II RIPs, through the studies of the recombinant Korean mistletoe lectin. To this end, we expressed, purified the recombinant Korean mistletoe lectin (rKML), and investigated its molecular characteristics in vitro, its cytotoxicity and ability to induce apoptotic cell death in cancer cells. To gain structural basis for its catalytic activity and sugar binding properties, we performed homology modeling studies based on the high degree of sequence identity and conserved secondary structure prediction between Korean and European, Himalayan mistletoe lectins, and Ricin.
Expression and Characterization of RNA-dependent RNA Polymerase of Dendrolimus punctatus Tetravirus
Zhou, Liang ; Zhang, Jiamin ; Wang, Xiaochun ; Jiang, Hong ; Yi, Fuming ; Hu, Yuanyang ;
BMB Reports , volume 39, issue 5, 2006, Pages 571~577
DOI : 10.5483/BMBRep.2006.39.5.571
Dendrolimus punctatus tetravirus (DpTV) has been identified as a new member of the genus Omegatetravirus of the family Tetraviridae that may be related serologically to Nudaurelia capensis virus (
). To establish the function of DpTV RNA genome and to better understand the mechanism of viral replication, the putative RNA-dependent RNA polymerase (RdRp) domain has been cloned and expressed in Escherichia coli. The recombinant protein was purified on a Ni-chelating HisTrap affinity column and demonstrated to initiate viral RNA synthesis in a primer-independent manner but not by terminal nucleotidyle transferase activity in the presence of
and RNA template. Mutation of the GDD to GAA interferes with the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive.
cDNA Cloning, Expression and Homology Modeling of a Luciferase from the Firefly Lampyroidea maculata
Emamzadeh, Abdo Rahman ; Hosseinkhani, Saman ; Sadeghizadeh, Majid ; Nikkhah, Maryam ; Chaichi, Mohammad Javad ; Mortazavi, Mojtaba ;
BMB Reports , volume 39, issue 5, 2006, Pages 578~585
DOI : 10.5483/BMBRep.2006.39.5.578
The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMP-binding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.
Germinal Center-independent Affinity Maturation in Tumor Necrosis Factor Receptor 1-deficient Mice
Kim, Jin-Ho ; Kim, Ju ; Jang, Yong-Suk ; Chung, Gook-Hyun ;
BMB Reports , volume 39, issue 5, 2006, Pages 586~594
DOI : 10.5483/BMBRep.2006.39.5.586
Germinal centers (GCs) have been identified as site at which the somatic mutation of immunoglobulins occurs. However, somatic mutations in immunoglobulins have also been observed in animals that normally do not harbor germinal centers. This clearly indicates that somatic mutations can occur in the absence of germinal centers. We therefore attempted to determine whether or not GCs exist in TNFR1-deficient mice, and are essential for the somatic mutation of immunoglobulins, using (4-hydroxy-3-nitropheny)acetyl-ovalbumin (NP-OVA). Both wild-type and TNFR1-deficient mice were immunized with NPOVA, and then examined with regard to the existence of GCs. No typical B-cell follicles were detected in the TNFR1-deficient mice. Cell proliferation was detected throughout all splenic tissue types, and no in vivo immune-complex retention was observed in the TNFR1-deficient mice. All of these data strongly suggest that no GCs were formed in the TNFR1-deficient mice. Although TNFR1-deficient mice are unable to form GCs, serological analyses indicated that affinity maturation had been achieved in both the wild-type and TNFR1-deficient mice. We therefore isolated and sequenced several DNA clones from wild-type and the TNFR1-deficient mice. Eight out of 12 wild-type clones, and 11 out of 14 clones of the TNFR-1-deficient mice contained mutations at the CDR1 site. Thus, the wild-type and TNFR1-deficient mice were not extremely different with regard to types and rates of somatic mutation. Also, high-affinity antibodies were detected in both types of mice. Collectively, our data appear to show that affinity maturation may occur in TNFR1-deficient mice, which completely lack GCs.
Molecular Analyses of the Metallothionein Gene Family in Rice (Oryza sativa L.)
Zhou, Gongke ; Xu, Yufeng ; Li, Ji ; Yang, Lingyan ; Liu, Jin-Yuan ;
BMB Reports , volume 39, issue 5, 2006, Pages 595~606
DOI : 10.5483/BMBRep.2006.39.5.595
Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.
Molecular Cloning and Characterization of a Novel Calcium-dependent Protein Kinase Gene IiCPK2 Responsive to Polyploidy from Tetraploid Isatis indigotica
Lu, Beibei ; Ding, Ruxian ; Zhang, Lei ; Yu, Xiaojing ; Huang, Beibei ; Chen, Wansheng ;
BMB Reports , volume 39, issue 5, 2006, Pages 607~617
DOI : 10.5483/BMBRep.2006.39.5.607
A novel calcium-dependent protein kinase gene (designated as IiCPK2) was cloned from tetraploid Isatis indigotica. The full-length cDNA of IiCPK2 was 2585 bp long with an open reading frame (ORF) of 1878 bp encoding a polypeptide of 625 amino acid residues. The predicted IiCPK2 polypeptide included three domains: a kinase domain, a junction domain (or autoinhibitory region), and a C-terminal calmodulin-like domain (or calcium-binding domain), which presented a typical structure of plant CDPKs. Further analysis of IiCPK2 genomic DNA revealed that it contained 7 exons, 6 introns and the length of most exons was highly conserved. Semi-quantitative RT-PCR revealed that the expression of IiCPK2 in root, stem and leaf were much higher in tetraploid sample than that in diploid progenitor. Further expression analysis revealed that gibberellin (
), NaCl and cold treatments could up-regulate the IiCPK2 transcription. All our findings suggest that IiCPK2 might participate in the cold, high salinity and GA3 responsive pathways.
The Enhanced Monocyte Adhesiveness after UVB Exposure Requires ROS and NF-κB Signaling in Human Keratinocyte
Park, Lee-Jin ; Ju, Sung-Mi ; Song, Ha-Yong ; Lee, Ji-Ae ; Yang, Mi-Young ; Kang, Young-Hee ; Kwon, Hyung-Joo ; Kim, Tae-Yoon ; Choi, Soo-Young ; Park, Jin-Seu ;
BMB Reports , volume 39, issue 5, 2006, Pages 618~625
DOI : 10.5483/BMBRep.2006.39.5.618
The infiltration of both monocyte and activated T cells in the skin is one of critical steps in the development of UVB-induced inflammation. Upregulation of adhesion molecules such as intercellular adhesion molecule 1 (ICAM-1) on the surface of keratinocytes plays an important role in this process. In this study, we examined the molecular mechanism responsible for UVB-induced expression of ICAM-1 and subsequent monocyte adhesion by keratinocyte. We observed that (1) UVB induced protein and mRNA expression of ICAM-1 in a dose- and time-dependent manner in human keratinocyte cell HaCaT; (2) UVB induced the translocation of NF-kappaB and inhibition of NF-kappaB by NF-kappaB inhibitors suppressed UVB-induced mRNA and protein expression of ICAM-1; (3) UVB increased the intracellular level of reactive oxygen species (ROS) by HaCaT cells; (4) UVB-induced increase of intracellular ROS level was suppressed by pre-treatment with diphenyl iodonium (DPI) and N-acetyl cysteine (NAC); and (5) inhibition of UVB-induced ROS production by DPI or NAC suppressed UVB-mediated translocation of NF-kappaB, expression of ICAM-1 and subsequent monocyte adhesion in HaCaT cells. These results suggest that UVB-induced ROS is involved in the translocation of NF-kappaB which is responsible for expression of ICAM-1 and subsequent increased monocyte adhesion in human keratinocyte.
Identification of a Novel Human Lysophosphatidic Acid Acyltransferase, LPAAT-theta, Which Activates mTOR Pathway
Tang, Wenwen ; Yuan, Jian ; Chen, Xinya ; Gu, Xiuting ; Luo, Kuntian ; Li, Jie ; Wan, Bo ; Wang, Yingli ; Yu, Long ;
BMB Reports , volume 39, issue 5, 2006, Pages 626~635
DOI : 10.5483/BMBRep.2006.39.5.626
Lysophosphatidic acid acyltransferase (LPAAT) is an intrinsic membrane protein that catalyzes the synthesis of phosphatidic acid (PA) from lysophosphatidic acid (LPA). It is well known that LPAAT is involved in lipid biosynthesis, while its role in tumour progression has been of emerging interest in the last few years. To date, seven members of the LPAAT gene family have been found in human. Here we report a novel LPAAT member, designated as LPAAT-theta, which was 2728 base pairs in length and contained an open reading frame (ORF) encoding 434 amino acids. The LPAAT-theta gene consisted of 12 exons and 11 introns, and mapped to chromosome 4q21.23. LPAAT-theta was ubiquitously expressed in 18 human tissues by RT-PCR analysis. Subcellular localization of LPAAT-theta-EGFP fusion protein revealed that LPAAT-theta was distributed primarily in the endoplasmic reticulum (ER) of COS-7 cells. Furthermore, we found that the overexpression of LPAAT-theta can induce mTOR-dependent p70S6K phosphorylation on Thr389 and 4EBP1 phosphorylation on Ser65 in HEK293T cells.
Interaction of Native and Apo-carbonic Anhydrase with Hydrophobic Adsorbents: A Comparative Structure-function Study
Salemi, Zahra ; Hosseinkhani, Saman ; Ranjbar, Bijan ; Nemat-Gorgani, Mohsen ;
BMB Reports , volume 39, issue 5, 2006, Pages 636~641
DOI : 10.5483/BMBRep.2006.39.5.636
Our previous studies indicated that native carbonic anhydrase does not interact with hydrophobic adsorbents and that it acquires this ability upon denaturation. In the present study, an apo form of the enzyme was prepared by removal of zinc and a comparative study was performed on some characteristic features of the apo and native forms by far- and near-UV circular dichroism (CD), intrinsic fluorescent spectroscopy, 1-anilino naphthalene-8-sulfonate (ANS) binding, fluorescence quenching by acrylamide, and Tm measurement. Results indicate that protein flexibility is enhanced and the hydrophobic sites become more exposed upon conversion to the apo form. Accordingly, the apo structure showed a greater affinity for interaction with hydrophobic adsorbents as compared with the native structure. As observed for the native enzyme, heat denaturation of the apo form promoted interaction with alkyl residues present on the adsorbents and, by cooling followed by addition of zinc, catalytically-active immobilized preparations were obtained.
Expression, Purification and Transduction of PEP-1-Botulinum Neurotoxin Type A (PEP-1-BoNT/A) into Skin
Kim, Dae-Won ; Kim, So-Young ; An, Jae-Jin ; Lee, Sun-Hwa ; Jang, Sang-Ho ; Won, Moo-Ho ; Kang, Tae-Cheon ; Chung, Kwang-Hoe ; Jung, Hyun-Ho ; Cho, Sung-Woo ; Choi, Jin-Hi ; Park, Jin-Seu ; Eum, Won-Sik ; Choi, Soo-Young ;
BMB Reports , volume 39, issue 5, 2006, Pages 642~647
DOI : 10.5483/BMBRep.2006.39.5.642
Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.