Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The gene encoding a 51 kDa polypeptide of Porphyromonas gingivalis 381 was isolated by immunoblotting using an antiserum raised against P. gingivalis alkaline phosphatase. DNA sequence analysis of a 2.5 kb DNA fragment containing a gene encoding the 51 kDa protein revealed one complete and two incomplete ORFs. Database searches using the FASTA program revealed significant homology between the P. gingivalis 51 kDa protein and the MurC protein of Escherichia coli, which functions in peptidoglycan synthesis. The cloned 51 kDa protein encoded a functional product that complemented an E. coli murC mutant. Moreover, the ORF just upstream of murC coded for a protein that was 31% homologous with the E. coli MurG protein. The ORF just downstream of murC coded for a protein that was 17% homologous with the Streptococcus pneumoniae penicillin-binding protein 2B (PBP2B), which functions in peptidoglycan synthesis and is responsible for antibiotic resistance. These results suggest that P. gingivalis contains a homologue of the E. coli peptidoglycan synthesis gene murC and indicate the possibility of a cluster of genes responsible for cell division and cell growth, as in the E. coli mra region.
...
PMID:A murC gene in Porphyromonas gingivalis 381. 749 15

We have delineated three short open reading frames, psiA, ORF-P and psiB within the psi operon of Rhizobium leguminosarum biovar phaseoli. psiA, in a multi-copy plasmid, causes inhibition of exopolysaccharide synthesis in R. leguminosarum. In addition, the suppression of exopolysaccharide synthesis due to the multi-copy psiA caused R. leguminosarum strains to stain with the dye calcofluor, a response that does not occur with wild-type strains of this species. Insertions of a defective phoA gene (lacking its promoter, ribosomal binding site and leader sequence) into psiA and psiB were isolated and the precise locations of the insertions were established. PsiA-PhoA and PsiB-PhoA protein fusions were found to express alkaline phosphatase activity indicating that PsiA and PsiB span the inner membrane or are translocated across it.
...
PMID:The psi operon of Rhizobium leguminosarum biovar phaseoli: identification of two genes whose products are located at the bacterial cell surface. 751 67

A secreted phosphodiesterase/alkaline phosphatase, APaseD, was purified from a culture of Bacillus subtilis JH646MS. Its phosphodiesterase activity was reminiscent of an APase isolated and characterized previously. Immunoassay and N-terminal sequencing showed the two proteins to be identical. Using the first 20 amino acids of the mature protein, a BLAST search of GenBank was used to find an homologous sequence. An exact match was found but in a putative non-coding region. It was hypothesized that there was a base pair deletion in the phoD gene. A DNA fragment internal to the coding region was generated by PCR using template DNA from a strain which produced APaseD. The PCR fragment was cloned and used to interrupt the gene. Western blot analysis of the parent and the mutated strains showed that APaseD was missing in the mutant. Resequencing of the gene revealed a larger ORF encoding a protein similar in size to the 49 kDa APaseD estimated by SDS-PAGE. The promoter was then cloned, sequenced and used in phoD-lacZ promoter fusions which showed that the gene was phosphate-starvation-induced and dependent on PhoP and PhoR for expression.
...
PMID:A Bacillus subtilis secreted phosphodiesterase/alkaline phosphatase is the product of a Pho regulon gene, phoD. 876 Sep 16

TnphoA mutagenesis of a Salmonella choleraesuis isolate recovered from septicemic infection of feeder pigs resulted in 56 PhoA+ KnR StrR mutants. Thirty-five mutants exhibited reduced levels of invasion in the Hep-2 cell model and were examined by SDS-PAGE Western Blot analysis using an anti-alkaline phosphatase antibody to visualize the insertion gene products. A mutant which produced a gene fusion product of 95 kDa and exhibited > 90% reduction in invasion was subcloned. A 10 Kb BamHI fragment of the chromosome containing the phoA insert was detected by hybridization and cloned into a pGEM vector. The resulting 1657 base sequence contained a 1104 bp ORF with two short regions of homology with S. typhimurium invF and invG. one region of homology with lcrD of Yersinia pseudotuberculosis but contained largely unique sequences not contained in Gene Bank. The full length sequence was not obtained as there was no stop codon detected. The % G+C was 44%, considerably lower than that of the Salmonella chromosome, but compatible with the proposed Yersinia origin of the inv genes. The NH2 387 a.a. sequence includes 5 transmembrane regions, resembling the model derived from the hydrophobicity plot of S. typhimurium InvA.
...
PMID:Unique Salmonella choleraesuis surface protein affecting invasiveness. Possible inv related sequence. 919 39

The nucleotide sequence of 17.3 kbp downstream of addA (98 degrees) on the Bacillus subtilis chromosome was determined. Twenty putative ORFs were identified. Three of them coincided with known B. subtilis genes, addA, sbcD and wprA. The product of four other ORFs showed similarity to SbcC of Clostridium perfringens, CotH of B. subtilis, 2-hydroxyhepta-2,4-diene-1,7-diodate isomerase of Methanococcus jannaschi and a putative ORF of Pseudomonas syringae. In addition, a sequence of 7.6 kbp downstream of citG (189 degrees) was analysed. Among 10 putative ORFs identified, two coincided with known genes, citG and mrgA, whilst three showed homology with X86780, a sensory protein kinase of Streptomyces hygroscopicus, an alkaline phosphatase regulatory protein and a hypothetical protease, YyxA, of B. subtilis.
...
PMID:Sequencing of regions downstream of addA (98 degrees) and citG (289 degrees) in Bacillus subtilis. 935 31

Chrysanthemyl diphosphate synthase (CPPase) catalyzes the condensation of two molecules of dimethylallyl diphosphate to produce chrysanthemyl diphosphate (CPP), a monoterpene with a non-head-to-tail or irregular c1'-2-3 linkage between isoprenoid units. Irregular monoterpenes are common in Chrysanthemum cinerariaefolium and related members of the Asteraceae family. In C. cinerariaefolium, CPP is an intermediate in the biosynthesis of the pyrethrin ester insecticides. CPPase was purified from immature chrysanthemum flowers, and the N terminus of the protein was sequenced. A C. cinerariaefolium lambda cDNA library was screened by using degenerate oligonucleotide probes based on the amino acid sequence to identify a CPPase clone that encoded a 45-kDa preprotein. The first 50 aa of the ORF constitute a putative plastidial targeting sequence. Recombinant CPPase bearing an N-terminal polyhistidine affinity tag in place of the targeting sequence was purified to homogeneity from an overproducing Escherichia coli strain by Ni(2+) chromatography. Incubation of recombinant CPPase with dimethylallyl diphosphate produced CPP. The diphosphate ester was hydrolyzed by alkaline phosphatase, and the resulting monoterpene alcohol was analyzed by GC/MS to confirm its structure. The amino acid sequence of CPPase aligns closely with that of the chain elongation prenyltransferase farnesyl diphosphate synthase rather than squalene synthase or phytoene synthase, which catalyze c1'-2-3 cyclopropanation reactions similar to the CPPase reaction.
...
PMID:Chrysanthemyl diphosphate synthase: isolation of the gene and characterization of the recombinant non-head-to-tail monoterpene synthase from Chrysanthemum cinerariaefolium. 1128 53

Brucella, an aerobic, nonsporeforming, nonmotile Gram-negative coccobacillus, is a NIH/CDC category B bioterror threat agent that causes incapacitating human illness. Medical defense against the bioterror threat posed by Brucella would be strengthened by development of a human vaccine and improved diagnostic tests. Central to advancement of these goals is discovery of bacterial constituents that are immunogenic or antigenic for humans. Outer membrane proteins (OMPs) are particularly attractive for this purpose. In this study, we cloned, expressed, and purified seven predicted OMPs of Brucella suis. The recombinant proteins were fused with 6-His and V5 epitope tags at their C termini to facilitate detection and purification. The B. suis surface genes were PCR synthesized based on their ORF sequences and directly cloned into an entry vector. The recombinant entry constructs were propagated in TOP 10 cells, recombined into a destination vector, pET-DEST42, then transformed into Escherichia coli BL21 cells for IPTG-induced protein expression. The expressed recombinant proteins were confirmed with Western blot analysis using anti-6-His antibody conjugated with alkaline phosphatase. These B. suis OMPs were captured and purified using a HisGrab plate. The purified recombinant proteins were examined for their binding activity with antiserum. Serum derived from a rabbit immunized intramuscularly with dialyzed cell lysate of Brucella rough mutant WRR51. The OMPs were screened using the rabbit antiserum and purified IgG. The results suggested that recombinant B. suis OMPs were successfully cloned, expressed and purified. Some of the expressed OMPs showed high binding activity with immunized rabbit antiserum.
...
PMID:Cloning, expression, and purification of Brucella suis outer membrane proteins. 1572 81

An exported 22 kDa putative lipoprotein was identified in an alkaline phosphatase gene fusion library of Mycobacterium avium subsp. paratuberculosis and expressed in Mycobacterium smegmatis. The full nucleic acid sequence of the gene encoding P22 was determined and the ORF was cloned into a mycobacterial expression vector, enabling full-length P22 to be produced as a C-terminal polyhistidine-tagged protein in M. smegmatis. N-terminal sequencing of the recombinant protein confirmed cleavage of a signal sequence. Native P22 was detected in culture supernatants and cell sonicates of M. avium subsp. paratuberculosis strain 316F using rabbit antibody raised to recombinant P22. Investigation of the presence of similar genes in other mycobacterial species revealed that the gene was present in Mycobacterium avium subsp. avium and similar genes existed in Mycobacterium intracellulare and Mycobacterium scrofulaceum. Database searches showed that P22 belonged to the LppX/LprAFG family of mycobacterial lipoproteins also found in Mycobacterium leprae and in members of the Mycobacterium tuberculosis complex. P22 shared less than 75% identity to these proteins. Recombinant P22 was able to elicit interferon-gamma secretion in blood from eight of a group of nine sheep vaccinated with a live attenuated strain of M. avium subsp. paratuberculosis (strain 316F) compared to none from a group of five unvaccinated sheep. Antibody to P22 was detected by Western blot analysis in 10 out of 11 vaccinated sheep, in two out of two clinically affected cows and in 11 out of 13 subclinically infected cows.
...
PMID:Identification and characterization of an immunogenic 22 kDa exported protein of Mycobacterium avium subspecies paratuberculosis. 1619 41

IkappaB kinase (IKK) play central roles in cell signaling by regulating nuclear factor-kappaB (NF-kappaB) activation, which is involved in inflammatory response, proliferation, development and bone homeostasis. We report here for the first time that an IKK homologue was cloned and functionally characterized in pearl oyster, Pinctada fucata. The full-length cDNA consists of 2546bp with an ORF encoding a 737 amino acids protein. The putative pearl oyster IKK protein (Pf-IKK) possesses the characteristic organization of the mammalian IKK proteins, namely an amino-terminal kinase domain followed by a leucine zipper region and a carboxylterminal helix-loop-helix motif. Real-time PCR (RT-PCR) analysis indicated that Pf-IKK was ubiquitously expressed in pearl oyster. We also found that lipopolysaccharides (LPS) transiently stimulates IkappaBalpha degradation, but not expression levels of Pf-IKK. When transfected into NIH3T3 cells, Pf-IKK activated the expression of NF-kappaB-controlled reporter gene and induced NF-kappaB translocation, whereas the activation was greatly deduced by pyrrolidine dithiocarbamate (PDTC). We also found that overexpression of Pf-IKK increased the alkaline phosphatase (ALP) activity significantly. Based on the results and the homology to the vertebrate NF-kappaB cascade, these studies help to highlight a potentially important regulatory pathway to the study of the related functions in mollusks.
...
PMID:Cloning and characterization of an IKK homologue from pearl oyster, Pinctada fucata. 1756 71

The Rab GTPases are important components of endocytic network in plant cells. Endocytosis participates in the cell's reaction to extracellular stimuli by desensitizing, down-regulating or recycling receptors and membrane proteins. Rab7 is a small GTP-binding protein involved in intracellular vesicle trafficking from late endosome to the vacuole. We have isolated Rab7 cDNA from Pennisetum glaucum, a relatively drought-stress tolerant food grain crop grown commonly in India, during cDNA-subtractive hybridization of dehydration-stress treated plants. The PgRab7 ORF, encoding 207 aminoacids, was over-expressed in E. coli. The recombinant PgRab7 protein showed GTP-binding and GTPase activity. Transcript expression of PgRab7 gene was differentially up-regulated by different environmental stimuli such as cold, dehydration and NaCl and also by a plant hormone IAA. Overexpression of PgRab7 gene enhanced tolerance to NaCl and mannitol in transgenic tobacco. Transgenic plants also had increased alkaline phosphatase (ALP) activity. These results show that PgRab7 is a potential candidate gene for developing both salinity and dehydration tolerance in planta.
...
PMID:Constitutive overexpression of a stress-inducible small GTP-binding protein PgRab7 from Pennisetum glaucum enhances abiotic stress tolerance in transgenic tobacco. 1789 98


1 2 Next >>

---