Bacillus anthracis grows in chains of rod-shaped cells, a trait that contributes to its escape from phagocytic clearance in host tissues. Using a genetic approach to search for determinants of B. anthracis chain length, we identified mutants with insertional lesions in secA2. All isolated secA2 mutants exhibited exaggerated chain length, ...

In recent years proteomics has come of age with the development of efficient tools for purification, identification and characterization of gene products predicted by genome projects. The intestinal protozoan Giardia intestinalis can be transfected but there is only a limited set of vectors available and most of them are not ...

While protein purification has long been dominated by standard chromatography, the relatively high cost and complex scale-up have promoted the development of alternative non-chromatographic separation methods. Here we developed a new non-chromatographic affinity method for the purification of proteins expressed in Escherichia coli. The approach is to genetically fuse the ...

Aim:  To evaluate an inter-generic recombinant alpha domain fusion protein for simultaneous detection and neutralization of Clostridium perfringens and Staphylococcus aureus alpha toxins. Methods and Results:  Truncated portions of Clostridial and Staphylococcal alpha hemolysin genes were PCR amplified and linked to each other through a hydrophilic flexible Glycine linker sequence ...

Prions, infectious agents causing transmissible spongiform encephalopathies (TSEs), are composed primarily of the pathogenic form (PrP(Sc) ) of the host-encoded prion protein (PrP(C) ). The susceptibility of sheep to scrapie is determined by polymorphisms in the coding region of the prion protein gene (PRNP), mainly at codons 136, 154, and ...

The high level expression of recombinant proteins in Escherichia coli often leads to the formation of inclusion bodies that contain most of the expressed protein held together by non-covalent forces. The inclusion bodies are usually solubilized using strong denaturing agents like urea and guanidium hydrochloride. In this study recombinant Omp28 ...

Bacillus subtilis lacking the SpoVT protein that regulates gene expression in developing forespores gave spores that released their dipicolinic acid (DPA) via germinant receptor (GR)-dependent germination more rapidly than wild-type spores. Non-GR germination via dodecylamine was more rapid with spoVT spores but Ca-DPA germination was slower. The effects of a ...

Amyloid-like fibrils from bovine whey protein have potential as efficient thickening and gelling agents for food and biomedical applications, but the link between fibril morphology and bulk viscosity is poorly understood. We examined how lyophilisation and rehydration affects the morphology and rheological properties of semiflexible (i.e. straight) and highly flexible ...

The actin cytoskeleton is one of the major targets of bacterial protein toxins. The family of binary actin-ADP-ribosylating toxins, including Clostridium difficile transferase CDT, Clostridiumperfringens iota toxin and Clostridiumbotulinum C2 toxin, modifies arginine-177 of actin. Thereby actin polymerization is blocked. By contrast, actin polymerization is facilitated by the tripartite Photorhabdus ...

Leishmania, Similarly to other eukaryotes, contains large amounts of actin and a number of actin related and binding proteins. Our earlier studies have shown that deletion of the gene corresponding to Leishmania actin depolymerizing protein (ADF/cofilin) adversely affects the flagellum assembly, intracellular trafficking and cell division. To further analyse this ...

Determining the localization, binding partners, and secondary modifications of individual proteins is crucial for understanding protein function. Several tags have been constructed for protein localization or purification under either native or denaturing conditions, but few tags permit all three simultaneously. Here, we describe a multifunctional tandem affinity purification (MAP) method ...

ABSTRACT: The mycotoxin T-2 toxin and Salmonella Typhimurium infections pose a significant threat to human and animal health. Interactions between both agents may result in a different outcome of the infection. Therefore, the aim of the presented study was to investigate the effects of low and relevant concentrations of T-2 ...

Directed evolution is a broadly successful strategy for protein engineering in the quest to enhance the stereoselectivity, activity, and thermostability of enzymes. To increase the efficiency of directed evolution based on iterative saturation mutagenesis, the adaptive substituent reordering algorithm (ASRA) is introduced here as an alternative to traditional quantitative structure-activity ...

Functionalized magnetic particles are emerging as a reliable and convenient technique in the purification of biomacromolecules (proteins and nucleic acids) and cell separation. In this study, we used novel solid nickel ferromagnetic particles coated with Protein A for the affinity purification of antibody. The study demonstrated that IgG can be ...

End Binding protein 1 (EB1) and Cytoplasmic Linker Protein of 170 kDa (CLIP-170) are two well-studied microtubule plus end-tracking proteins (+TIPs) that target to growing microtubule plus ends in the form of comet tails and regulate microtubule dynamics. However, the mechanism by which they regulate microtubule dynamics is not well ...

In this manuscript we describe the two-step purification of a mono-PEGylated anti-epidermal growth factor receptor (EGFR) single-chain Fv. A weak cation exchanger was used for capture. Elution using arginine suppressed protein aggregation and allowed a very good resolution with purity and product-recovery was above 90%. Free PEG was removed completely. ...

We investigated the vascular transport properties of exogenously applied proteins to Brassica oleracea plants and compared their delivery to various aerial parts of the plant with carboxy fluorescein (CF) dye. We identified unique properties for each protein. Alexafluor-tagged bovine serum albumin (Alexa-BSA) and Alexafluor-tagged Histone H1 (Alexa-Histone) moved slower than ...

Multimodular polyketide synthases (PKSs) have an assembly line architecture in which a set of protein domains, known as a module, participates in one round of polyketide chain elongation and associated chemical modifications, after which the growing chain is translocated to the next PKS module. The ability to rationally reprogram these ...

Asparagine-linked glycosylation is an essential and highly conserved protein modification reaction. In eukaryotes, oligosaccharyl transferase (OT), a multisubunit membrane-associated enzyme complex, catalyzes this reaction in newly synthesized proteins. In Saccharomyces cerevisiae, OT consists of nine non-identical membrane proteins. Ost4p, the smallest subunit, bridges the catalytic subunit Stt3p with Ost3p. Mutation ...

Flagellin has the ability to polymerize into long filaments under appropriate conditions. Our work aims at the construction of flagellin-based fusion proteins which possess polymerization ability and preserve the functional properties of the fusion partner as well. The hypervariable D3 domain of Salmonella flagellin, containing residues 190-283, is a good ...

In this study, the lipoxygenase (ana-LOX) gene from Anabaena sp. PCC 7120 was successful expressed and secreted in Bacillus subtilis. Under the control of the P43 promoter, with a signal peptide from the B. subtilis 168 nprB gene, and facilitated by the molecular chaperone PrsA, the production of the recombinant ...

A synthetic de novo designed heterodimeric coiled-coil was used to copurify two target fluorescent proteins, Venus and enhanced cyan fluorescent protein (ECFP). The coiled-coil consists of two 21-amino acid repetitive sequences, (EIAALEK)(3) and (KIAALKE)(3) , named E3 and K3, respectively. These sequences were fused to the C-termini of ECFP or ...

The large scale production of a monoclonal anti-progesterone antibody in serum free medium followed by affinity chromatography on protein G lead to a contamination of the antibody sample with a protein of about 14kDa. This protein was identified by mass spectrometry as secretory leukocyte protease inhibitor (SLPI). This SLPI contamination ...

Protein fusion tags are indispensible tools used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Solubility-enhancing tags, genetically engineered epitopes, and recombinant endoproteases have resulted in a versatile array of combinatorial elements that facilitate protein detection and purification in microbial ...

Due to the high costs associated with purification of recombinant proteins the protocols need to be rationalized. For high-throughput efforts there is a demand for general methods that do not require target protein specific optimization(1) . To achieve this, purification tags that genetically can be fused to the gene of ...

A point mutation in Prnp that converts tyrosine (Y) at position 145 into a stop codon leading to a truncated prion molecule as found in an inherited transmissible spongiform encephalopathy (TSE), Gertsmann-Sträussler-Scheincker syndrome, suggests that the N-terminus of the molecule (spanning amino acids 23-144) likely plays a critical role in ...

The production of membrane proteins in the large quantities necessary for structural analysis requires many optimization steps. The GFP-fusion-based scheme described in earlier chapters ( Chapters 4 , 8 , and 16 ) facilitates these steps by allowing the selection of high yielding clones that produce detergent-stable membrane proteins. Here, ...

A point mutation in Prnp that converts tyrosine (Y) at position 145 into a stop codon leading to a truncated prion molecule as found in an inherited transmissible spongiform encephalopathy (TSE), Gertsmann-Sträussler-Scheincker syndrome, suggests that the N-terminus of the molecule (spanning amino acids 23-144) likely plays a critical role in ...

Transcription efficiency of inducible promoters remains a bottleneck in recombinant protein production in Bacillus subtilis cells. Here, we present experimental data how to generate strong IPTG-inducible promoters by optimization of nucleotides at the conserved regions of the groESL promoter including the UP element, the -35, -15, -10 and the +1 ...

The application of surface modified magnetic adsorbent particles in combination with magnetic separation techniques has received considerable awareness in recent years. There is a particular need in protein purification and analysis for specific, functional and generic methods of protein binding on solid supports. Nanoscale superparamagnetic iron oxide particles have been ...

Two fluorescent proteins that emit in the far-red and infrared range for imaging applications in cells and in vivo are described.

Spermatids generate diverse and unusual actin and microtubule populations during spermiogenesis to fulfill mechanical and cargo transport functions assisted by motor and non-motor proteins. Disruption of cargo transport may lead to teratozoospermia and consequent male infertility. How motor and non-motor proteins utilize the cytoskeleton to transport cargos during sperm development ...

Recently, wheat gluten has been proposed as technological adjuvant in order to clarify wines. However, the possibility that residual gluten proteins remain in treated wines cannot be excluded, representing a hazard for wheat allergic or celiac disease patients. In this work, commercial wheat glutens, in both partially hydrolyzed (GBS-P51) and ...

Organisms limited by carbon, nitrogen or sulphur can reduce protein production costs by transitions to less costly amino acids, or by reducing protein expression. These alternative mechanisms of nutrient thrift might respond differently to selection, but this possibility remains untested. We hypothesized that relatively invariant sequence composition responds to long-term ...

Protein-losing enteropathy (PLE) is a rare complication of a variety of intestinal disorders characterized by an excessive loss of proteins into the gastrointestinal tract due to impaired integrity of the mucosa. The clinical presentation of patients with PLE is highly variable, depending upon the underlying cause, but mainly consists of ...

OBJECTIVE: To study the role and significance of the polycomb group (PcG) protein EZH2 (enhancer of zeste homolog 2) in the multi-step process of intestinal-type gastric carcinogenesis. METHODS: Gastric specimens were obtained from 142 patients with gastric disease, including 34 with chronic non-atrophic gastritis (NCAG), 33 chronic atrophic gastritis (CAG) ...

A 24-year-old man presented with abdominal distension, diarrhea, and nausea. Blood tests showed eosinophilia (WBC 14400/microl, Eos 36%) and slight hypoproteinemia (TP 6.4 mg/dl, Alb 3.7 mg/dl). Ultrasonography and computed tomography revealed massive ascites (WBC 11500/microl, Eos 95%, protein 4.7 g/dl) and wall thickening of the small intestine. Endoscopic and ...

We report a 25-year-old Malay man with Subutex-related endocarditis, complicated by protein-losing enteropathy from severe tricuspid regurgitation and congestive heart failure. The intestinal protein loss was reversed with surgical valvular intervention. This case highlights the importance of recognising the rare association between protein-losing enteropathy and congestive heart failure in the ...

Protein-losing enteropathy (PLE) is defined as a condition in which excess protein loss into the gastrointestinal lumen is severe enough to produce hypoproteinaemia. Previously reported procedures for the detection of protein loss have many limitations (such as rapid reabsorption of the radiolabel, unstable protein binding both in vivo and in ...

Intestinal lymphangiectasia is a rare condition, which is characterized by the dilation of small bowel lymphatics and presents with signs and symptoms of protein-losing enteropathy. Some patients have complained of occlusive symptoms attributable to the mechanical obstruction caused by the considerable mucosal edema associated with the lymphatic dilation. On the ...

Protein-losing enteropathy (PLE) is an uncommon manifestation associated with systemic lupus erythematosus (SLE). Here, a case with SLE and concomitant hypoalbuminemia is reported. Technetium-99m albumin scintigraphy demonstrated a localized lesion in the ascending colon, and the diagnosis of SLE-related PLE was established. Due to a poor response to medical treatment, ...

Protein-losing enteropathy (PLE) is characterized by gastrointestinal loss of serum protein. It is usually caused by hypersecretion from a tumor, ulcer, or long standing lymphangiectasia. However, we report a 47-year-old man of peritoneal nodal follicular lymphoma who developed PLE with none of them. Oozing of whitish fluid from duodenal bulbar ...

Protein-losing enteropathy may uncommonly complicate cardiac disease. While well described as a complication for patients having undergone previous Fontan surgery for congenital heart disease, pericardial and valvular aetiologies are much less frequent. We report a 35-year-old female presenting with marked hypoalbuminaemia and peripheral oedema on a background of known rheumatic ...

A 17-year-old boy was admitted to the hospital twice in a year for 2 episodes of hemolytic uremic syndrome (HUS). During these 2 HUS episodes he had diarrhea, decreased serum complement, decreased total protein and decreased serum albumin concentrations. We suggest that protein-losing enteropathy and hypocomplementemia due to intestinal lymphangiectasia ...

We report an unusual case of systemic lupus erythematosus presented with protein-losing enteropathy. A 24-year-old girl was referred to our hospital with generalized edema, thrombocytopenia, hypoalbuminemia, hypercholesterolemia, hypocomplementemia, antinuclear antibody (ANA) (speckled pattern) and anti- SSA/Ro positivities, and elevated CA125 antigen appeared in the blood examination. On the radiological studies, ...

Gastrointestinal bleeding has not been reported in patients with protein-losing enteropathy. The investigators describe 3 patients with protein-losing enteropathy and gastrointestinal bleeding after Fontan operations. Bleeding occurred at the onset of protein-losing enteropathy, was macroscopic, led to anemia, and required blood transfusion in all patients. Invasive testing failed to demonstrate ...

Protein-losing enteropathy (PLE), the loss of plasma proteins through the intestine, is a symptom in ostensibly unrelated diseases. Emerging commonalities indicate that genetic insufficiencies predispose for PLE and environmental insults, e.g. viral infections and inflammation, trigger PLE onset. The specific loss of heparan sulfate (HS) from the basolateral surface of ...

We report the clinical, laboratory, endoscopic and histopathological findings in a 40-year-old woman with watery diarrhoea and hypoproteinaemia. Elevated alpha(1)-antitrypsin clearance confirmed massive protein-losing enteropathy. Gastroscopic and colonoscopic biopsies showed abundant infiltration of the small bowel wall with eosinophils in proximal duodenum and terminal ileum, respectively. These findings established the ...

Insecticidal activity of proteinous venom from tentacle of jellyfish Rhopilema esculentum Kishinouye was determined against three pest species, Stephanitis pyri Fabriciusa, Aphis medicaginis Koch, and Myzus persicae Sulzer. R. esculentum full proteinous venom had different insecticidal activity against S. pyri Fabriciusa, A. medicaginis Koch, and M. persicae Sulzer. The 48 ...

Constrictive pericarditis represents a rare cause of protein-losing enteropathy due to intestinal lymphangiectasia. We report the case of a patient with an atypical clinical presentation of constrictive pericarditis and protein-losing enteropathy as its principal manifestation; he was successfully treated with pericardiectomy. We conclude that, constrictive pericarditis should be considered in ...