Electronic Thesis and Dissertation Repository

Degree

Master of Science

Program

Biochemistry

Supervisor

Dr. Shaw

Abstract

Parkin is an E3 ubiquitin ligase which degrades misfolded proteins and prevents the formation of abnormal protein aggregates often formed in Parkinson’s disease. The main goal of this thesis was to perform structural analysis on the IBR(In-Between-RING)-RING2 (Really Interesting New Gene) domain of parkin. After determining the three-dimensional solution structure of the protein by NMR spectroscopy, the RING2 domain was identified to be similar to the IBR domain, showing that it is not a canonical RING domain. The catalytic cysteine on RING2 was also shown to be solvent exposed, supporting the recently proposed RING/HECT hybrid mechanism of parkin as an RBR E3 ligase. The structure also revealed that IBR and RING2 domains do not interact. This was confirmed with two dimensional NMR experiments and split GFP system. The 14 disease-state IBR-RING2 proteins were analyzed using NMR spectroscopy to monitor the structural impact of autosomal recessive juvenile parkinsonism (ARJP) related mutations.

Included in

Biochemistry Commons

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