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Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 21 for “"Sortase"”.
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Applications of Sortase A from Staphylococcus aureus
Sortase A (SrtA) mediated ligation provides a mild and site-specific method for attaching a wide range of molecular probes to biological molecules, such as proteins and DNA. This method does not require any toxic reagents or harsh conditions to achieve high level of labelling, and the product could …
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Identifying Sortase A Variants With Higher Catalytic Effeciency
… studies because of its substrate specificity. Sortase-mediated transpeptidation is one approach that has been well documented. Staphylococcus aureus sortase A (SrtAstaph), a membrane-anchored cysteine transpeptidase present in gram-positive bacteria, covalently anchors virulence-associated …
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Sortase as a Tool in Biotechnology and Medicine
… harnessed two reactions catalyzed by the enzyme sortase A and applied them to generate new methods for the purification and site-selective modification of recombinant protein therapeutics. </p><p>We utilized native peptide ligation —a well-known function of sortase A— to attach a small molecule …
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SORTASE AND CHEMICAL BIOLOGY AS TOOLS TO STUDY UBIQUITINATION
… in this field, we leverage the power of sortase, a bacterial transpeptidase enzyme crucial to cell wall synthesis, to develop new strategies for studying protein ubiquitination. As one new technology, we introduce point mutations to the ubiquitin sequence in combination with click …
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Sortase-mediated ubiquitylation of Histone H2B and its biological consequences
… H2B molecule that could be used for intranuclear sortase-mediated histone ubiquitylation. For the benefit of future SrtA experiments, we produced mouse polyclonal antibodies against Staphylococcus aureus and Streptomyces pyogenes SrtA. These experiments will help to further the development of …
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Site-specific protein labeling via sortase A and its applications/
… synthesized probes. This system exploits sortases, a class of bacterial transpeptidases, that recognize a small five amino acid tag genetically fused to the protein of interest and catalyze the formation of an amide bond between the protein to be studied and the probe. The second part of …
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Regulation of Surface Proteins Assembly on the Wall of Gram-Positive Bacteria
… through a transpeptidation reaction, whereby sortase cleaves a conserved Cterminal LPXTG motif and attaches the protein to the peptidoglycan precursor lipid II. This thesis deals with the spatial regulation of the sorting reaction, and the role sortase localization plays in the correct …
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Sortases: Keystones to Virulence and Targets for Anti-Infective Therapy
… class of anti-virulence chemotherapeutics is the sortase class of enzymes. Sortases are extracellular transpeptidases unique to Gram-positive bacteria. Their function is to covalently attach secreted virulence factors to the bacterial cell wall. Deletion or inhibition of sortases results in severe …
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Development of VHH- and antibody- based imaging and diagnostic tools
… Chemoenzymatic labeling of molecules using sortase allows site-specific attachment of a single label of interest to the target protein containing the sortase recognition sequence LPXTG. VHHs specific for immune cell determinants labeled with sortase technology facilitate non-invasive and …
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Engineered red blood cells and their applications
… This system employs genetic engineering and sortase A, a bacterial transpeptidases.
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Molecular Basis of Corynebacterium Diphtheriae Virulence and Infection In The Caenorhabditis Elegans Model Host
… by a specific transpeptidase enzyme called sortase. Although the SpaA pili are shown to be specific for pharyngeal cells in vitro, little is known about functions of the three pili in bacterial pathogenesis. This is mainly due to lack of in vivo models of corynebacterial infection. As an …
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Mechanisms of bacterial CRISPR immunity and a phage counter-defense
… protein subunits within a complex using a sortase-mediated labeling strategy (Chapter 3). Finally, we determine how the anti-CRISPR protein AcrIIA11 inhibits diverse Cas9 orthologs (Chapter 4).
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Enzymatic crosslinking of dynamic hydrogels for in vitro cell culture
… using enzymes with orthogonal reactivity. Sortase A (SrtA) and mushroom tyrosinase (MT) were used sequentially to achieve initial gelation and on-demand stiffening. In addition, hydrogels permitting reversible stiffening through SrtA-mediated peptide ligation were established. Specifically, …
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Targeting the tight junction : immunotherapy of colon cancer
… protein drugs is presented. Finally, the use of sortase A as a means to generate fusion proteins posttranslationally is described. Taken together, this additional work demonstrates a productive intersection of basic research and protein engineering methods.
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Construction of plasmid-based expression and secretion vectors and study of the immobilization of proteins on the surface of Bacillus subtilis cells
… bacteria code for one or more enzymes termed sortases that catalyze the covalent anchoring of substrate proteins on their cell wall. They recognize an amino acid sequence designated sorting motif, present close to the C-terminal end of the substrate proteins, cleave within this motif, and …
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Análises in silico das proteínas de superfície e secretadas de Staphylococcus aureus para discriminar entre as mastites bovina clínica e subclínica
… nos genes clfa (fator de aglutinação A), srta (sortase A) e sspa (protease). Uma proteína transportadora (cl3316) foi identificada in silico como exclusiva das cepas subclínicas. Diferentes programas de predição foram utilizados para a identificação das proteínas de superfície e secretadas das …
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Delivery of biomolecules into mammalian cells using anthrax toxin
… pore. A facile, modular ligation strategy using sortase A was developed for the conjugation of biomolecules to LFN. The biomolecules for this analysis included antibody mimic proteins with defined sizes and secondary structures, mirror image peptides and proteins, polypeptides containing …
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A 3D synthetic extracellular matrix that can be dissolved to interrogate the cellular local microenvironment on demand
… from the 3D matrix by repurposing a mutant of sortase A (SrtA). We demonstrate SrtA-mediated gel dissolution preserves important cell morphological features and can be used to measure recovered cytokines much more accurately compared to other proteolytic degradation methods. We observe …
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Development and Application of Mass Spectrometry Based Methodologies to Study Ubiquitin-Like Modifiers
… of the bacterial protease- transpeptidase Sortase A (SrtA). Using a mutant with increased catalytic activity, SrtA mediates biotinylation of diglycine tryptic remnants, and also acts as a specific protease for release of isopeptides from streptavidin for analysis by mass spectrometry. This …
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Recording Cellular Interactions in Germinal Centers and Beyond
… uLIPSTIC eliminates this limitation by anchoring Sortase A (SrtA) and its oligoglycine acceptor non-specifically to the cell membrane at high density. This innovation allows enzymatic labeling between any membranes that come into close proximity, vastly expanding the range of potential …
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