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Query: EC:2.7.7.49 (
reverse transcriptase
)
31,746
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
A galactose-specific dimeric
lectin
from pinto beans was purified using a procedure that involved affinity chromatography on Affi-gel blue gel, anion exchange chromatography on Q-Sepharose, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 200. The molecular mass of this homodimeric
lectin
was 62 kDa and that of each of its subunits was 31 kDa. The hemagglutinating activity of pinto bean
lectin
was stable within the pH range of 3-12 and the temperature range of 0-70 degrees C. By using the [3H-methyl]-thymidine incorporation assay, it was shown that the
lectin
had the ability to evoke a mitogenic response from murine splenocytes but it did not inhibit proliferation of L1210 leukemia cells. The pinto bean
lectin
inhibited HIV-1
reverse transcriptase
with an IC50 of 3 microM.
...
PMID:Purification and characterization of a galactose-specific lectin with mitogenic activity from pinto beans. 1660 May 11
A
lectin
, with a molecular mass of 79 kDa, and with specificity toward rhamnose and O-nitrophenyl-beta-D-galactopyranoside, was isolated from samta tomato fruits. The procedure entailed ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose and gel filtration by fast protein liquid chromatography on Superdex 75. The
lectin
was unadsorbed on DEAE-cellulose but adsorbed on Affi-gel blue gel and CM-cellulose. The
lectin
was stable up to 70 degrees C. The
lectin
activity was potentiated by NaOH solutions (25-100 mM), but was reduced by 50 and 100 mM HCl solutions. The activity of the
lectin
was reduced in the presence of CaCl(2), MgCl(2) and ZnCl(2), but was potentiated by 5 and 10 mM AlCl(3) solutions. The
lectin
stimulated the mitogenic response in mouse splenocytes and inhibited human immunodeficiency virus-1
reverse transcriptase
with an IC(50) of 6.2 microM.
...
PMID:A lectin with some unique characteristics from the samta tomato. 1677 25
We previously generated a double knockout mutant (act/aopB) of a diarrheal isolate SSU of A. hydrophila, in which the genes encoding Aeromonas outer membrane protein B (AopB), a structural component of the type III secretion system (T3SS), and a type II (T2)-secreted cytotoxic enterotoxin gene (act) were deleted. This mutant exhibited minimal virulence in mice, compared to animals infected with wild-type (WT) A. hydrophila. Based on microarray analyses, WT A. hydrophila altered the expression of 434 and 80 genes in murine macrophages (RAW 264.7) and human colonic epithelial cells (HT-29), respectively. Approximately half of these gene expression alterations were abrogated when host cells were infected instead with the act/aopB mutant. In this study, we used microarrays to examine early host transcriptional responses in spleens of mice infected for 3h with WT A. hydrophila or its act/aopB mutant. Our data indicated that expression of 221 genes was altered (158 up-regulated and 63 down-regulated) in spleens of WT bacteria-infected animals. There were 21 genes that were consistently more highly expressed in WT A. hydrophila-infected mice, compared to mice infected with its act/aopB mutant. Ten of these genes were either induced to a lesser extent (e.g., interleukin-6, macrophage inflammatory protein-2, and cyclooxygenase-2), not altered at all (e.g., killer cell
lectin
-like receptor subfamily B member A), or down-regulated (e.g., cytochrome P450) in animals infected with A. hydrophila, compared to phosphate-buffered saline-infected control animals, when the mutant was used instead of the WT. We verified the microarray results at the transcript level by performing real-time
reverse transcriptase
-polymerase chain reaction on selected genes and at the protein level by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. This is the first study demonstrating in vivo gene regulation in mice infected with A. hydrophila and the contribution of virulence factors and host responses to the disease process.
...
PMID:Global transcriptional responses of wild-type Aeromonas hydrophila and its virulence-deficient mutant in a murine model of infection. 1736 24
NKG2D is an activating
lectin
-like receptor that initiates natural killer (NK) cell responses against transformed tumor cells expressing its ligands, i.e., molecules related to major histocompatibility complex (MHC) class I molecules. NKG2D lacks signaling elements in its cytoplasmic domain and can deliver stimulatory signals only in association with transmembrane adaptor proteins DAP10 or DAP12. The complementary DNAs (cDNAs) encoding the bovine homologues of NKG2D and the adaptor proteins DAP10 and DAP12 were cloned by
reverse transcriptase
-polymerase chain reaction (RT-PCR) from resting bovine peripheral blood mononuclear cells (PBMC) and sequenced. Comparison with human, pig, and mouse sequences showed that bovine NKG2D is most similar to pig NKG2D and short mouse NKG2D (NKG2D-S). Similar to its human, mouse, and pig homologues, the cDNA for bovine DAP10 codes for a phosphatidyl-inositol-3 (PI-3) kinase-binding site (YxxM) in its cytoplasmic region. Finally, similar to its human, mouse, and pig homologues, the cDNA encoding bovine DAP12 demonstrates one tyrosine-based activated motif (ITAM) in its cytoplasmic domain. Bovine NKG2D cell surface expression was analyzed by flow cytometry on HEK 293 cells transiently transfected with cDNA expression vectors encoding COOH-terminal polyhistidine-tagged NKG2D and NH(2)-terminal Flag-tagged DAP10 and DAP12. Confirming previous findings for short mouse NKG2D-S, bovine NKG2D immunoreceptor could associate with either DAP10 or DAP12 adaptor protein for its cell surface expression.
...
PMID:Cloning, sequencing, and cell surface expression pattern of bovine immunoreceptor NKG2D and adaptor molecules DAP10 and DAP12. 1753 Feb 42
In the present work, we followed a microarray approach to analyze the expression of glycosylation-related genes on different cell populations obtained from mouse thymus. Among other genes, transcription of the two-domain type galectin-8 was detected both in thymocytes and thymic epithelial cells (TECs), which was confirmed by
reverse transcriptase
(RT)-PCR assays independently carried out on both cell populations. Two splice variants, differing solely in the presence of a nine amino acid insertion in the linker peptide region connecting the two carbohydrate recognition domains (CRDs), were identified from purified thymocytes. Expression of galectin-8 was verified at the protein level in total organ extracts by western-blots of lactosyl-Sepharose purified binders. To explore the possible biological roles of locally produced galectin-8, both splice variants were recombinantly expressed in bacteria and assayed over cultured thymocytes. In spite of their binding to all cell populations, addition of either isoform of galectin-8 to thymocyte cultures induced apoptosis only of the CD4(high)CD8(high) cells through caspases pathway activation. All of these effects were prevented by the addition of thiodigalactoside (TDG) or lactose, thus indicating that the proapoptotic activity of galectin-8 was due to the specific interaction of its CRDs with defined cell surface glycans. Together, our results demonstrate intrathymic expression of galectin-8 in mouse, and suggest an active role for this
lectin
in shaping the mature T cell repertoire.
...
PMID:Galectin-8 induces apoptosis in the CD4(high)CD8(high) thymocyte subpopulation. 1789 94
The objective of the present study was to isolate a
lectin
from fresh fruiting bodies of the mushroom Pleurotus citrinopileatus and examine it for various biological activities. The isolation procedure comprised ion exchange chromatography on DEAE-cellulose, CM-celluloses, and Q-Sepharose, and gel filtration on Superdex 75. A homodimeric 32.4 kDa
lectin
displaying high hemagglutinating activity was isolated with over 110 fold of purification. Its N-terminal amino acid sequence, QYSQMAQVME, has not been reported for other lectins. The
lectin
was unadsorbed on DEAE-cellulose in 0.001 M NH4HCO3 buffer (pH 9.4), but adsorbed on CM-cellulose in 0.001 M NH4OAc buffer (pH 4.8) and eluted by approximately 0.05 M NaCl in the same buffer. The
lectin
was subsequently adsorbed on Q-Sepharose and eluted by a linear gradient of 0-0.2 M NaCl in 10 mM NH4HCO3 buffer (pH 8.5). The
lectin
was obtained in a purified form after gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of the
lectin
was inhibited by maltose, O-nitrophenyl-beta-d-galactopyranoside, O/P-nitrophenyl-beta-d-glucuronide and insulin. It was stable at temperatures up to 60 degrees C, and in NaOH and HCl solutions up to 0.1 M and 0.006 M concentration, respectively. It was sensitive to inhibition by HgCl2 and potentiation by AlCl3. The
lectin
exerted potent antitumor activity in mice bearing sarcoma 180, and caused approximately 80% inhibition of tumor growth when administered intraperitonealy at 5 mg/kg daily for 20 days. It elicited a mitogenic response from murine splenocytes in vitro with the maximal response at a
lectin
concentration of 2 microM. The
lectin
inhibited HIV-1
reverse transcriptase
with an IC50 of 0.93 microM. It was devoid of antifungal activity.
...
PMID:A novel lectin with potent antitumor, mitogenic and HIV-1 reverse transcriptase inhibitory activities from the edible mushroom Pleurotus citrinopileatus. 1796 26
It has been believed that clonal propagation by asexual reproduction has serious disadvantages for long-term survival, because asexual reproduction seems not to remove harmful mutations, it seems not to give rise to genetic variations upon which evolution depends and it seems not to reset cell aging. In this article, we re-consider those arguments, by reviewing asexual reproduction of the tunicate, Polyandrocarpa misakiensis. Tracer experiments of bud formation and growth using morphological and chimeric phenotypes showed that the parental epithelial tissues surrounding the bud primordium do not enter the growing bud. It is possible, therefore, to assume that budding involves the purge of a large number of parental somatic cells and tissues. Unlike sexuals, asexuals do not carry out meiotic recombination nor gene shuffling that are two major sources of genetic variation, but we can show that in P. misakiensis at least two genes have significant redundancy and genetic variation even in a clonal colony. Telomerase expressed in germlines is thought to reset the molecular clock executed by telomere shortening. In our Polyandrocarpa cDNA projects, four out of about 2,000 cDNAs examined were matched with retroviral
reverse transcriptase
that is the catalytic subunit of telomerase, suggesting that telomerase might work in asexual reproduction. In P. misakiensis, dedifferentiation system is used to make new asexual generations. TC14
lectin
plays an important role in the maintenance of multipotent but differentiated state of the formative tissue. It is antagonized by tunicate serine protease (TRAMP) that has striking mitogenic and dedifferentiation-inducing activities on the multipotent cells. This system would serve to delay aging of somatic cells. In conclusion, empirical arguments that asexual reproduction is disadvantageous to long-term life do not appear to be tenable to budding of P. misakiensis.
...
PMID:Advantage or Disadvantage: Is Asexual Reproduction Beneficial to Survival of the Tunicate, Polyandrocarpa misakiensis. 1849 80
Oxidative stress induced by superoxide plays an important role in pathogenesis of cardiovascular diseases. NAD(P)H oxidase is a principal enzymatic origin for superoxide in vasculature. Recently, novel homologues of cytosolic components of NAD(P)H oxidase, Nox organizer 1 (NOXO1) and Nox activator 1 (NOXA1), are identified. On the other hand, oxidized low-density lipoprotein (ox-LDL) generates reactive oxygen species (ROS) in endothelial cells via
lectin
-like oxidized low-density lipoprotein receptor-1 (LOX-1). In the present investigation, the authors examined the expression, the regulation, and the role of NOXA1 in the generation of ROS in endothelial cells. The expression of NOXA1 was confirmed by
reverse transcriptase
-polymerase chain reaction (RT-PCR). Dihydroethidium method showed that ox-LDL and angiotensin II increased the generation of intracellular ROS. Once the expression of p22(phox) or NOXA1 was suppressed by siRNA, the generation of ROS induced by ox-LDL and angiotensin II were potently decreased. Moreover, the expression of NOXA1 was increased by ox-LDL in a time-and dose-dependent manner. In conclusion, endothelial NOXA1 plays an essential role in generation of ROS. Ox-LDL not only increased the generation of ROS via LOX-1, but also enhanced the expression of NOXA1 in endothelial cells. NOXA1 is likely a key player that links ox-LDL with the activation of endothelial NAD(P)H oxidase.
...
PMID:Essential role of NOXA1 in generation of reactive oxygen species induced by oxidized low-density lipoprotein in human vascular endothelial cells. 1856 54
A mannose/glucose-specific
lectin
has been purified from Chinese evergreen chinkapin (Castanopsis chinensis) seeds, one of the most popular foods in East Asia. This
lectin
, designated as CCL, exhibited hemagglutinating activity in mouse and rabbit erythrocytes. It displayed a single band with a molecular mass of 29 kDa in SDS-PAGE and a 120-kDa peak in gel-filtration on Superdex-200. Its hemagglutinating activity was stable in the pH range 6-12 and at temperatures below 60 degrees C. The N-terminal amino acid sequence of CCL differed from those of other lectins in the same family. CCL inhibited the proliferation of HepG2 cells and adult emergence in fruitflies. CCL exhibited mitogenic activity toward mouse splenocytes, and induced nitric oxide production from mouse peritoneal macrophages but was devoid of inhibitory activity toward mycelial growth and HIV-1
reverse transcriptase
.
...
PMID:A mannose/glucose-specific lectin from Chinese evergreen chinkapin (Castanopsis chinensis). 1857 Aug 98
When Crassostrea gigas oyster sperm acrosome react a ring of bindin protein is exposed that bonds the sperm to the egg vitelline envelope. The putative functional unit of bindin is a fucose
lectin
(F-lectin) domain that is structurally conserved among phyla. There is only one bindin gene in C. gigas, which can possess 1-5 tandem F-
lectin
repeats. Alternative splicing can alter the number of repeats per bindin mRNA. Recombination occurs in a highly variable intron in the middle of each F-
lectin
repeat to create many different
lectin
domain sequences [Moy, G.W., Springer, S.A., Adams, S.L., Swanson, W.J., Vacquier, V.D., 2008. Extraordinary intraspecific diversity in oyster sperm bindin. Proc. Natl. Acad. Sci. U.S.A. 105, 1993-1998]. Two bindin genes were sequenced to learn more about bindin introns. The first gene (6914 bp) contained one F-
lectin
repeat. The second gene (25,932 bp) contained three tandem F-
lectin
repeats. Four of the introns in this larger gene are conserved in size among individuals. However, the one intron in each F-
lectin
repeat is highly variable in size and sequence, indicating that it has been a hot spot for recombination. A retroposon with high
reverse transcriptase
homology is present in the three repeat gene immediately upstream of the first F-
lectin
repeat, suggesting that retroposition is one mechanism by which F-
lectin
repeats are duplicated. The retroposon is not present in the one F-
lectin
repeat bindin gene. Three GA microsatellites, one in each intron immediately upstream of the start of each F-
lectin
repeat exon, and one downstream CT microsatellite, suggest that loopout strand hybridization can occur, and
lectin
repeats replicate and transpose within the gene. The CT microsatellite is not found in the one F-
lectin
repeat containing gene. Oysters appear to use every possible mechanism to create variation in the F-
lectin
domains of sperm bindin. This is presumably in response to sexual conflict that operates in the prevention of polyspermy.
...
PMID:Bindin genes of the Pacific oyster Crassostrea gigas. 1867 29
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