Unraveling the mechanism of multicopper oxidases : from ensemble to single molecule

Gupta, A. (2014)

Doctoral Thesis

Ever since the structural data of biological macromolecules became available, there has been consistent struggle to relate this new information to the existing spectroscopy, activity and theoretical descriptions of these proteins and to understand the evolution and/or to predict the role of yet uncharacterized gene products in this light. The research presented in this thesis primarily deals with understanding the structure__function relationship of a newly discovered blue copper protein. The protein is derived from Streptomyces coelicolor and is called small laccase (SLAC). It utilizes four copper ions to catalyze the oxidation of substrate molecules concomitant with the reduction of oxygen to water. The catalytic cycle of this enzyme is studied using a variety of spectroscopic and kinetic methods in an attempt to improve our understanding of the internal operations which are critical to its functioning. The new results obtained are presented in this thesis.

• EPR
• Enzyme kinetics
• Single molecule spectroscopy
• FRET
• Electron transfer
• Oxidoreductase

• All authors: Gupta, A.
• Supervisor: Canters, G.W.; Aartsma, T.J.
• Committee: Eliel, E.R.; Groenen, E.J.J.; Farver, O.; Scrutton, N.S.
• Qualification: Doctor (dr.)
• Awarding Institution: Leiden Institute of Physics (LION), Faculty of Science, Leiden University
• Date: 2014-04-29
• ISBN (print): 9789085931829