Dynamin 2 (DNM2) is a large GTPase involved in the release of nascent vesicles during endocytosis and intracellular membrane trafficking. Distinct DNM2 mutations, affecting middle domain (MD) Pleckstrin homology (PH), have been identified autosomal dominant centronuclear myopathy (CNM) intermediate axonal forms Charcot-Marie-Tooth peripheral neuropathy (CMT). We report here first CNM mutation (c.1948G>A, p.E650 K) effector (GED), leading to slowly progressive moderate myopathy. COS7 cells...

Summary Background Cow's milk allergy (CMA) is one of the most widespread human allergies, especially in young children. Although CMA intensively studied, little known about recognition patterns allergens allergic patients, and determination these a prerequisite for development efficient diagnostic prognostic tools. Several factors present difficulties such determination, because (i) contains large number potential allergens; (ii) majority consist complex suspensions rather than solutions;...

A cluster of arginine biosynthetic genes Corynebacterium glutamicum ATCC 13032, comprising argJ, argB and argD as well part argC argF, has been cloned by heterologous complementation an Escherichia coli argE mutant. The gene order established argCJBDF sequencing the entire 4.4 kb DNA fragment. C. can be transcribed in E. cells from internal promoter located coding preceding argJ gene, whereas transcription appears vector-dependent. Expression corynebacterial is repressed native host but not...

A genomic DNA fragment encoding aminoacylase activity of the eubacterium Bacillus stearothermophilus was cloned into Escherichia coli. Transformants expressing were selected by their ability to complement E. coli mutants defective in acetylornithine deacetylase activity, enzyme that converts N-acetylornithine ornithine arginine biosynthetic pathway. The 2.3-kb has been entirely sequenced. Analysis sequence revealed two open reading frames, one which encoded aminoacylase. B. aminoacylase,...

A 3·4 kb EcoRI fragment, cloned in E. coli, that carries part of a cluster genes encoding arginine biosynthetic functions the thermophilic bacterium Bacillus stearothermophilus, was sequenced on both strands. The sequence consists truncated argC gene, an argJ region polypeptide with N-acetylglutamate synthase and ornithine acetyltransferase activities, argB gene N-terminal argD. encodes 258-amino-acid deduced M r 26918. very high thermostable 5-phosphotransferase activity detected extracts...

Summary In recent years, natural pigments in living organisms have attracted increased interest as an excellent biological material for the development of new biotechnological approaches. Melanin, a large polymeric pigment, has important functions cells, including free radical scavenging and radiation protection, making this molecule attractive medical nutritional purposes. The variety forms melanin requires use various approaches to its purification. study, we developed simple yet unique...

The argJ gene coding for N 2 ‐acetyl‐ l ‐ornithine: ‐glutamate ‐acetyltransferase, the key enzyme involved in acetyl cycle of ‐arginine biosynthesis, has been cloned from thermophilic procaryotes: archaeon Methanoccocus jannaschii , and bacteria Thermotoga neapolitana Bacillus stearothermophilus . Archaeal only complements an Escherichia coli argE mutant (deficient acetylornithinase, which catalyzes fifth step linear biosynthetic pathway), whereas bacterial genes additionally complement argA...

We report here the cloning of arginine repressor gene argR Bacillus stearothermophilus and characterization purification to homogeneity its product. The deduced amino acid sequence 16.8‐kDa ArgR subunit shares 72% identity with mesophilic homologue AhrC Bacilus subtilis . Sequence analysis B. comparisons repressors suggest that thermostable comprises an N‐terminal DNA‐binding a C‐terminal oligomerization arginine‐binding region. has been overexpressed in E. coli purified as 48.0‐kDa trimeric...

Abstract The protein array methodology is used to study DNA‐protein and protein‐protein interactions governing gene expression from the Bacillus stearothermophilus P argCo promoter‐operator region. Using probes labelled with near‐infrared fluorescence dyes exitation characteristics close 700 or 800 nm, it possible detect signals proteins (purified non‐purified in Escherichia coli cell extracts) immobilised on a nitrocellulose membrane high sensitivity (almost 12 amol of spotted for...

Summary: The expression of Bacillus stearothermophilus genes complementing arginine auxotrophs Escherichia coli was studied. activity responsible for the formation ornithine in B. identified as a repressible acetyltransferase (genetic symbol argJ) encoded by same DNA fragment argC, orgA and argB genes. subtilis licheniformis displayed pattern enzyme activities stearothermophilus. In contrast to previous reports, these organisms consequently use cyclic pathway biosynthesis. also possesses...

Abstract The comprehensive and quantitative analysis of the protein phosphorylation patterns in different cellular context is considerable general interest. ability to quantify discrete signalling proteins large collections biological samples would greatly favour development systems biology field cell signalling. Reverse‐phase array (RPPA) potentially represents a very attractive approach map signal transduction networks with high throughput. In present report, we describe an improved...

Peptide ligand-induced dimerization of the extracellular region epidermal growth factor receptor (sEGFR) is central to signal transduction many cellular processes. A small molecule microarray screen has been developed search for non-peptide compounds able bind sEGFR. We describe discovery nitro-benzoxadiazole (NBD) that enhance tyrosine phosphorylation EGFR and thereby trigger downstream signaling pathways other kinases in cancer cells. The protein profile cells exposed NBD some extent...

Activation of cell signaling by reactive chemicals and pollutants is an important issue for human health. It has been shown that lipophilic nitro-benzoxadiazole (NBD) compounds rapidly move across the plasma membrane enhance Epidermal Growth Factor Receptor (EGFR) tyrosine phosphorylation in cancer cells. Unlike ligand-dependent activation, mechanism this induction relies on generation hydrogen peroxide, which involved activation catalytic site receptor inactivation protein phosphatase...

Large scale comparative evaluation of protein expression requires miniaturized techniques to provide sensitive and accurate measurements the abundance molecules present as individual and/or assembled complexes in cells. The principle competition between target for binding arrayed antibodies has recently been proposed assess differential numerous proteins with one-color or two-color fluorescence detection methods. To establish limiting factors validate use alternative profiling, we performed...

Bacterial promoters, which increase the efficiency of gene expression, differ from other promoters by several characteristics. This difference, not yet widely exploited in bioinformatics, looks promising for development relevant computational tools to search strong bacterial genomes. We describe a new triad pattern algorithm that predicts promoter candidates annotated genomes matching specific patterns group I σ70 factors Escherichia coli RNA polymerase. It detects promoter-specific motifs...

Abstract Porphyrins have a unique aromatic structure determining particular photochemical properties that make them promising photosensitizers for anticancer therapy. Previously, we synthesized set of artificial porphyrins by modifying side-chain functional groups and introducing different metals into the core structure. Here, performed comparative study binding 29 cationic with plasma proteins using microarray spectroscopic approaches. The were noncovalently immobilized onto...

The expression and activity of DNA-dependent protein kinase (DNA-PK) is related to DNA repair status in the response cells exogenous endogenous factors. Recent studies indicate that Epidermal Growth Factor Receptor (EGFR) involved modulating DNA-PK. It has been shown a compound 4-nitro-7-[(1-oxidopyridin-2-yl)sulfanyl]-2,1,3-benzoxadiazole (NSC), bearing nitro-benzoxadiazole (NBD) scaffold, enhances tyrosine phosphorylation EGFR triggers downstream signaling pathways. Here, we studied...

Protoplasts of the entomopathogenic bacterium Bacillus thuringiensis subsp. galleria were transformed by plasmid pBC16. The frequency transformation was much lower than that subtilis. All isolated B. transformants characterized increased sensitivity to lysozyme as compared with original strain.

Targeting epidermal growth factor receptor (EGFR) with tyrosine kinase inhibitors (TKI) has been widely exploited to disrupt aberrant phosphorylation flux in cancer. However, a bottleneck of potent TKIs is the acquisition drug resistance mutations, secondary effects, and low ability attenuate tumor progression. We have developed an alternative means targeting EGFR that relies on protein degradation through two consecutive routes, ultimately leading cancer cell detachment-related death....