Application of Electrospray Ionization Mass Spectrometry to Study Protein-Ligand Interactions and Enzyme Kinetics

• Author / Creator

  Rezaei Darestani, Reza

• This thesis describes the development and application of electrospray ionization mass spectrometry (ESI-MS) to study the noncovalent protein-ligand interactions and also investigate the kinetic of enzymatic reactions. The rate of glycolipids cleavage by the human neuraminidase 3 (hNEU3) was studied using time-resolved ESI-MS assay. The ESI-MS method was validated by comparing the relative hydrolysis rates of soluble glycolipids obtained from ESI-MS aasay with fluorescence-based assay. The kinetic analysis revealed that picodiscs present a better presentation of glycolipids for enzymatic studies. A catch-and-released electrospray ionization mass spectrometry (CaR-ESI-MS) assay was employed to screen a series of anti-cancer drugs against target protein. The bound ligands were then identified and relatively quantified using collision-induced dissociation (CID). The assay was capable of observing specific interactions between anti-cancer drugs and protein of interest.

• Subjects / Keywords

  • Protein-Ligand Interactions
  • Electrospray Ionization
  • Mass-spectrometry
  • Enzyme Kinetics

• Graduation date

  Fall 2015

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R3S757063

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.