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Biochemical and molecular characterization of putative immunoprotective molecules of the soft
Biochemical and molecular characterization of putative immunoprotective molecules of the soft

... peptides from hemolymph, such as defensins and lysozyme, while less is known about bacterial recognition molecules, or antimicrobial mechanisms in other tissues. The current study attempted to identify novel antimicrobial mechanisms, with a focus on bacterial recognition by hemolymph proteins and an ...
The bacterial translation stress response
The bacterial translation stress response

... (i.e. without involvement of EF-Tu) to a vacant ribosomal A-site if the corresponding aminoacylated species is unavailable, thereby stalling translation (Fig. 1b). RelA recognizes the arrested ribosomes and catalyzes the synthesis of (p)ppGpp. The synthesis of (p)ppGpp by RelA is proposed to reduce ...
BTXpred: Prediction of bacterial toxins
BTXpred: Prediction of bacterial toxins

... made for predicting the bacterial toxins and their types. One of the major problems was how to create datasets of bacterial toxins, endotoxin, exotoxins and different type of exotoxins, as there is no database available on bacterial toxins. It was beyond the scope of this study to classify the avail ...
Protein Signatures Distinctive of Alpha Proteobacteria and Its
Protein Signatures Distinctive of Alpha Proteobacteria and Its

... α-proteobacteria have been divided into seven main subgroups or orders (viz. Caulobacterales, Rhizobiales, Rhodobacterales, Rhodospirillales, Rickettsiales, Sphingomondales, and Parvularucales) (Maidak et al. 2001; Garrity & Holt 2001; Kersters et al. 2003). However, the branching order and interrel ...
Structure and Composition of Resistant Layers in Bacterial Spore
Structure and Composition of Resistant Layers in Bacterial Spore

... received little attention, even though resistance to enzymes must be an important attribute contributing to the survival of spores in natural environments. The results in this paper suggest that at least two reasonably well-defined spore components are important in determining their resistance to en ...
Filamentous hemagglutinin of bordetella pertussis as a carrier
Filamentous hemagglutinin of bordetella pertussis as a carrier

... humans. In addition, the use of the present methods and conjugates is not limited to prophylactic application; therapeutic application are also contemplated (e.g., AIDS therapy and post-exposure prophylaxis). A vaccine composition which can be useful in the vaccination against meningitis caused by H ...
Thesis - doktori
Thesis - doktori

... animals. BCG vaccine was first used in humans against tuberculosis in 1921. Since that time, BCG is the most widely used vaccine all over the world, but its protective value as anti-TB vaccine is questionable. BCG can protect children against severe forms of the disease, particularly meningitis, but ...
Flagellar glycosylation – a new component of the motility repertoire?
Flagellar glycosylation – a new component of the motility repertoire?

... noteworthy that a significant number of the flagellin structural protein monomers which are part of these alternative flagellar structures have recently been shown to be glycosylated at multiple sites (Table 1). As thousands of copies of these proteins are present within an assembled filament the bi ...
The Tricky Ways Bacteria Cope with Iron Limitation
The Tricky Ways Bacteria Cope with Iron Limitation

... FhuA was in vivo reconstituted from the b barrel domain and the plug domain, both equipped with a signal sequence for secretion across the cytoplasmic membrane (Braun et al. 2003a, b). The yield of active protein was rather high (45 %). This suggests that the b barrel folds independent of the plug a ...
Comparative and Functional Genomic Analysis of Prokaryotic Nickel
Comparative and Functional Genomic Analysis of Prokaryotic Nickel

... transport systems of several different types. We integrated in silico and in vivo methods for the analysis of various protein families containing both nickel and cobalt transport systems in prokaryotes. For functional annotation of genes, we used two comparative genomic approaches: identification of ...
2‐1 GENOMIC ANALYSIS REVEALS MULTIPLE [FeFe
2‐1 GENOMIC ANALYSIS REVEALS MULTIPLE [FeFe

... development of the concept of this energy rich gas being the central free intermediate in the conversion of plant biomass in wood feeding termites (56). ...
Sequencing, sub-cloning, expression and purification of
Sequencing, sub-cloning, expression and purification of

... bacterial genera comprising the sphingomonads have a unique capacity for degrading PAHs and thus have a promising future for bioremediation of these contaminants in the soil. Traditional methods of remediation involve costly excavation and incineration of contaminated soil and sediment. Bioremediati ...
LIN, a Novel Type of U-Box/WD40 Protein
LIN, a Novel Type of U-Box/WD40 Protein

... ENOD20, and ENOD40) were expressed in lin at comparable levels to the wild type, while late nodulins such as MtN6, ENOD2, and ENOD8 failed to be induced in lin. This suggests that LIN acts downstream of the early Nod factor signal transduction pathway ...


... Regulation of gene expression is an essential mechanism that allows bacteria to rapidly adapt to alterations in their environment. Gene expression in bacteria is mainly controlled at the level of transcription initiation. To achieve this control a number of different mechanisms have evolved, one of ...
Immunological Tests
Immunological Tests

... Immunological Tests ...
Interference of Staphylococcus aureus virulence factors in the blood
Interference of Staphylococcus aureus virulence factors in the blood

... from other staphylococcus genus, since S.aureus is the only one that produces this enzyme. Whether it is during primary infection or when the bacteria spreads to other tissues and forms abscesses, at some point during S. aureus infection, staphylococcal cells will enter the bloodstream (Lowy 1998). ...
MTSR is a Dual Regulator that Controls Virulence Genes and
MTSR is a Dual Regulator that Controls Virulence Genes and

... bound to heme-containing proteins such as hemoglobin, myoglobin, hemosiderin, and heme-type enzymes (Otto, Verweij-van Vught et al. 1992; Edison, Bajel et al. 2008). Hemoglobin, the most abundant metalloprotein in humans, carries more than two thirds of the total iron. Hemoglobin and heme that are r ...
No Slide Title
No Slide Title

... Mechanism of action: Interacts with the bacterial plasma membrane via the lipid portion to form a pore. Metabolic death occurs by leakage of electrolytes without lysis of the bacteria ...


... may playa role in this process have been studied. Whereas some attachments are achieved by non-fimbrial structural adhesins that are present as monomers or oligomers on the outer membrane, other attachment is mediated by surface organelles such as fimbriae or pili (Abraham et ai., 1998; O'Toole and ...
Molecules of Streptococcus gordonii that bind to
Molecules of Streptococcus gordonii that bind to

... a partial breakdown product that, under certain experimental conditions, is absent due to complete processing. Antibodies raised to the 62/60 proteins also reacted with the 45 kDa molecule. Thus, these molecules are antigenically related and may be breakdown products of a larger molecule of 170 kDa ...
Genome-sequence analysis of Acinetobacter johnsonii MB44
Genome-sequence analysis of Acinetobacter johnsonii MB44

... penicillin-binding protein 7/8. Moreover, one siderophore biosynthesis gene cluster and one capsular polysaccharide gene cluster, which were predicted to be important virulence factors for C. elegans, were identified in the MB44 genome. The current study demonstrated that A. johnsonii MB44, with its ...
Rb sphaeroides
Rb sphaeroides

... line) and without (dashed line) the pufX protein present. Inset: Magnification of the spectra in the accessory BChl and Bpheo region. From [2]. ...
Staphylococcus aureus
Staphylococcus aureus

... Workers at dairy plant were found recycling products returned by large-lot customers and unshipped products to make new batches. Cartons were opened manually with bare hands and poured into a storage vat for recycling. ...
Photoactivation mechanism of PAmCherry based on crystal
Photoactivation mechanism of PAmCherry based on crystal

... 4° (Fig. 1 A). As none of the torsion angles is 0° or 180°, the C␤ atom in Tyr-67 has sp3 hybridization, not sp2 hybridization as observed in mCherry (15). This suggests that PAmCherry1 in the OFF state has a single bond, not a double bond, between the C␤ atom in Tyr-67 and the imidazol-5-ol ring. T ...
Structure and functional characterization of pyruvate decarboxylase from Gluconacetobacter diazotrophicus
Structure and functional characterization of pyruvate decarboxylase from Gluconacetobacter diazotrophicus

... *Values in brackets indicate assay pH. #Calculated based on values given in reference [5]. ...
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Thermal shift assay



A thermal shift assay quantifies the change in thermal denaturation temperature of a protein under varying conditions. The differing conditions that can be examined are very diverse, e.g. pH, salts, additives, drugs, drug leads, oxidation/reduction, or mutations. The binding of low molecular weight ligands can increase the thermal stability of a protein, as described by Koshland (1958) and Linderstrom-Lang and Schellman (1959). Almost half of enzymes require a metal ion co-factor. Thermostable proteins are often more useful than their non-thermostable counterparts, e.g. DNA polymerase in the polymerase chain reaction, so protein engineering often includes addingmutations to increase thermal stability. Protein crystallisation is more successful for proteins with a higher melting point and adding buffer components that stabilise proteins improve the likelihood of protein crystals forming.If examining pH then the possible effects of the buffer molecule on thermal stability should be taken into account along with the fact that pKa of each buffer molecule changes uniquely with temperature. Additionally, any time a charged species is examined the effects of the counterion should be accounted for.Thermal stability of proteins has traditionally been investigated using biochemical assays, circular dichroism, or differential scanning calorimetry. Biochemical assays require a catalytic activity of the protein in question as well as a specific assay. Circular dichroism and differential scanning calorimetry both consume large amounts of protein and are low-throughput methods. The thermofluor assay was the first high-throughput thermal shift assay and its utility and limitations has spurred the invention of a plethora of alternate methods. Each method has its strengths and weaknesses but they all struggle with intrinsically disordered proteins without any clearly defined tertiary structure as the essence of a thermal shift assay is measuring the temperature at which a protein goes from well-defined structure to disorder.
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