Global ETD Search
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 12 of 12 for “"Single molecule pull down"”.
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Probing cellular protein complexes using single-molecule pull-down
… result from dynamic interactions between biomolecules. The gold standard method for investigating interaction between biomolecules is the pull-down assay. We have extended the conventional pull-down assay to its ultimate limit: the analysis of single biomolecular complexes. We achieve this …
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Single molecule pull-down of viral RNA-protein complexes
Protein and RNA molecules interact with multiple protein partners to perform essential cellular processes such as post-transcriptional regulation of mRNA. Recently, a single molecule pull-down (SiMPull) assay was developed to isolate and study single protein complexes directly from cell lysates. …
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Single molecule pull down of double strand RNA binding proteins
RNA molecules are transcribed as single stranded naturally, but with most of them forming in to structures composed of duplex regions, loop, bulge or mismatches. RNAs with double stranded regions, or known as double-strand RNAs (dsRNAs). The class of proteins responsible for processing dsRNAs is …
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Characterisation of neurodegenerative diseases derived protein aggregates using improved single-molecule pull-down
… this thesis begins by establishing an improved single-molecule pull-down (SiMPull) assay, which was used to characterise soluble α-Synuclein (αS) and Amyloid-β (Aβ) aggregates in serum from patients with PD and controls. Our findings reveal that the ratio of detected aggregates (αS/(Aβ+αS)) and …
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Single-molecule profiling of tau aggregates in tauopathies: From test tube to patient
In recent decades, single-molecule methods have transformed the physical sciences, offering unprecedented insights into the complexity of biological systems at molecular resolution. In particular, fluorescence-based methods now enable the detection and imaging of individual biomolecules and their …
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Single-molecule techniques for mapping protein aggregates
… are heterogeneous in sizes and structures, single-molecule techniques are required to unravel their compositions. Super-resolution microscopy is particularly useful, as these aggregates are smaller than the diffraction limit of light. Compared to conventional light microscopy, the …
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Characterisation and Detection of P53 Aggregates
… In this thesis, a series of advances in single-molecule assays are presented to characterise full-length p53 aggregates and detect p53 aggregates in human plasma. Firstly, a single-molecule fluorescence microscope with flat-field illumination was constructed and characterised. Secondly, a …
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Mechanical manipulation and visualization of single biomolecules using fluorescence spectroscopy
… resonance energy transfer) assay to interrogate single biomolecules under applied forces between ~ 0.3 and 28 pN. In human cells, the ends of chromosomes are capped by 100-200 nt long telomeric DNA, consisting of tandem hexanucleotide guanine (G)-rich repeats. Four of such telomeric, G-rich …
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Role of ORCA and ORC in chromatin organization and DNA replication
… Suv39H1 in a chromatin context-dependent manner. Single-molecule pull-down assays demonstrate that ORCA-ORC and multiple H3K9 KMTs exist in a single complex and that ORCA stabilizes H3K9 KMT complex. Cells lacking ORCA show alterations in chromatin architecture, with significantly reduced H3K9 di- …
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ORCA/LRWD1, a novel player in the initiation of eukaryotic DNA replication
… the stoichiometry of these associations via single-molecule pull-down experiments that each molecule of ORCA can bind to one molecule of ORC, one molecule of Cdt1, and two molecules of Geminin. Further, ORCA directly interacts with the N-terminus of Orc2, and this association is essential for …
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Redefining the specificity of phosphoinositide-binding by human PH domain-containing proteins
… shortcomings, our laboratory has developed a single-molecule pull down (SiMPull) assay to probe protein-lipid binding. This method, called lipid-SiMPull, has high specificity and sensitivity while utilizing lipid vesicles of desired composition and full-length proteins expressed in whole cell …
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Development of single-molecules techniques to study native α-Synuclein aggregates
… (<5% of a solution of protein). For this reason, single-molecule fluorescence microscopy has been used to study them; by probing molecules one-by-one, the rare oligomeric species can be identified amongst the monomeric protein, counted and characterised. Typically, to study species in …