Spelling suggestions: "subject:"biomolecular eecognition"" "subject:"biomolecular arecognition""
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Nanotechnology for Molecular Recognition of Biological AnalytesTriulzi, Robert C. 23 January 2009 (has links)
Nanotechnology is a term used to describe nanometer scaled systems. This thesis presents various nanomaterials and systems for the investigation of biologically relevant analytes in general, and in particular for their detection, decontamination, or destruction. The validation of short peptide fragments as models for protein aggregation is initially discussed through applying spectroscopic and microscopic techniques to Langmuir monolayer surface chemistry. Following this validation, the use of nanogold as a photoablative material for the destruction of aggregated protein is investigated. Subsequently, the versatility of nanotechnology is shown by investigating a different form of nanogold; namely, gold quantum dots and the interesting phenomenon that arise when dealing with materials on a nanoscale. Experiments involving a complex between these gold quantum dots and an antibody are performed for the detection of an immunoglobulin in solution. The power of this analytical technique is highlighted by the capability of detecting the analyte at nanomolar concentrations. Finally, a limitation-the multiple synthetic steps necessary for imparting biological activity-- of quantum dots is addressed: a single step reaction is studied that allows for direct stabilization and conjugation of quantum dots with proteins and enzymes. As a representative application of the above mentioned procedure, the detection and decontamination of an organophosphorus compound is explored. In general, methods for overcoming limitations of nanoparticles and nanocrystals are discussed.
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Conformational Stability!? : Synthesis and Conformational Studies of Unnatural Backbone Modified PeptidesNorgren, Anna S. January 2006 (has links)
<p>The beauty of the wide functionality of proteins and peptides in Nature is determined by their ability to adopt three-dimensional structures. This thesis describes artificial molecules developed to mimic secondary structures similar to those found crucial for biological activities.</p><p>In the first part of this thesis, we focused on post-translational modifications of a class of unnatural oligomers known as <i>β</i>-peptides. Through the design and synthesis of a glycosylated <i>β</i><sup>3</sup>-peptide, the first such hybrid conjugate was reported. In this first report, a rather unstable 3<sub>14</sub>-helical structure was found. Subsequently, a collection of six new glycosylated <i>β</i><sup>3</sup>-peptides was synthesized with the aim to optimize the helical stability in water.</p><p>The ability of natural proteins, i.e. lectins, to recognize the carbohydrate residue on these unnatural peptide backbones was investigated through a biomolecular recognition study.</p><p>The second part of this thesis concerns the design of conformationally homogeneous scaffolds, which could be of importance for biomedical applications. In paper V, four- and five-membered cyclic <i>all</i>-<i>β</i><sup>3</sup>-peptides were investigated for this purpose. In a subsequent paper, a completely different strategy was employed; herein, the ability of a single <i>β</i><sup>2</sup>-amino acid to restrict the conformational freedom of a cyclic α-peptide was studied. </p><p>In the third part of this thesis, we synthesized and investigated the folding propensities of novel backbone modified oligomers, i.e. <i>β</i>-peptoids (<i>N</i>-substituted <i>β</i>-Ala) with α-chiral side chains.</p><p>The collective results of these studies have established the procedures required for synthesis of glycosylated <i>β</i>-peptides and deepened our understanding of the factors governing folding among such oligomers. Moreover, it was established that <i>β</i>-amino acids can be a useful tool to increase conformational stability of cyclic peptides.</p>
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Conformational Stability!? : Synthesis and Conformational Studies of Unnatural Backbone Modified PeptidesNorgren, Anna S. January 2006 (has links)
The beauty of the wide functionality of proteins and peptides in Nature is determined by their ability to adopt three-dimensional structures. This thesis describes artificial molecules developed to mimic secondary structures similar to those found crucial for biological activities. In the first part of this thesis, we focused on post-translational modifications of a class of unnatural oligomers known as β-peptides. Through the design and synthesis of a glycosylated β3-peptide, the first such hybrid conjugate was reported. In this first report, a rather unstable 314-helical structure was found. Subsequently, a collection of six new glycosylated β3-peptides was synthesized with the aim to optimize the helical stability in water. The ability of natural proteins, i.e. lectins, to recognize the carbohydrate residue on these unnatural peptide backbones was investigated through a biomolecular recognition study. The second part of this thesis concerns the design of conformationally homogeneous scaffolds, which could be of importance for biomedical applications. In paper V, four- and five-membered cyclic all-β3-peptides were investigated for this purpose. In a subsequent paper, a completely different strategy was employed; herein, the ability of a single β2-amino acid to restrict the conformational freedom of a cyclic α-peptide was studied. In the third part of this thesis, we synthesized and investigated the folding propensities of novel backbone modified oligomers, i.e. β-peptoids (N-substituted β-Ala) with α-chiral side chains. The collective results of these studies have established the procedures required for synthesis of glycosylated β-peptides and deepened our understanding of the factors governing folding among such oligomers. Moreover, it was established that β-amino acids can be a useful tool to increase conformational stability of cyclic peptides.
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Synthetic phosphorylation of kinases for functional studies in vitroChooi, Kok Phin January 2014 (has links)
The activity of protein kinases is heavily dependent on the phosphorylation state of the protein. Kinase phosphorylation states have been prepared through biological or enzymatic means for biochemical evaluation, but the use of protein chemical modification as an investigative tool has not been addressed. By chemically reacting a genetically encoded cysteine, phosphocysteine was installed via dehydroalanine as a reactive intermediate. The installed phosphocysteine was intended as a surrogate to the naturally occurring phosphothreonine or phosphoserine of a phosphorylated protein kinase. Two model protein kinases were investigated on: MEK1 and p38α. The development of suitable protein variants and suitable reaction conditions on these two proteins is discussed in turn and in detail, resulting in p38α-pCys180 and MEK1-pCys222. Designed to be mimics of the naturally occurring p38α-pThr180 and MEK1-pSer222, these two chemically modified proteins were studied for their biological function. The core biological studies entailed the determination of enzymatic activity of both modified proteins, and included the necessary controls against their active counterparts. In addition, the studies on p38α-pCys180 also included a more detailed quantification of enzymatic activity, and the behaviour of this modified protein against known inhibitors of p38α was also investigated. Both modified proteins were shown to be enzymatically active and behave similarly to corresponding active species. The adaptation of mass spectrometry methods to handle the majority of project's analytical requirements, from monitoring chemical transformations to following enzyme kinetics was instrumental in making these studies feasible. The details of these technical developments are interwoven into the scientific discussion. Also included in this thesis is an introduction to the mechanism and function of protein kinases, and on the protein chemistry methods employed. The work is concluded with a projection of implications that this protein chemical modification technique has on kinase biomedical research.
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