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Investigation of Novel Bioorthogonal Chemical Reactions on Proteins and Amyloid Formation

Rezaei, Mohsen | 2017

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 49483 (03)
  4. University: Sharif University of Technology
  5. Department: Chemistry
  6. Advisor(s): Kalhor, Hamid Reza
  7. Abstract:
  8. Bioorthogonal chemistry defines any reaction, inside or outside the living systems, without interfering with native chemical processes and 3dimensional structure of proteins. In these types of reactions water is the sole solvent; a neutral pH is required; the temperature must be up to 40 oC. The kinetics of reactions must be on the hour scale and the nontoxic reagents with low concentration must be used. One of the simplest methods of visualizing protein molecule is covalent attachment of fluorescein to protein. In this work several fluorescent compounds were synthesized. These novel compounds include acylchloride fluorescein, N-hydroxysuccinimide fluorescein, thiophenol fluorescein, and gluconolactone fluorescein. O-methyl fluorescein and O-dimethyl fluorescein were also synthesized. O-dimethyl fluorescein was converted to a novel compound with yellow fluorescence. Subsequently, the fluorescent compounds were examined in following applications: 1) covalent attachment to protein, 2) the effect on amyloid (miss folding) formation, 3) activity based proteome profiling for enzyme that bind to inhibitors. The gel electrophoresis was employed to visualize covalent attachments of synthetic compounds to proteins. Labeled proteins were separated from small unbounded molecules via dialysis and size exclusion chromatography. In this work the synthesized compounds were identified with 1HNMR, UV and MS analysis
  9. Keywords:
  10. Enzymatic Inhibition ; Amyloid Inhibitors ; Bioorthogonal Chemistry ; Fluoresein ; Amyloid Formation ; Proteome Profiling

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