MSc Thesis Presentation: Amide proton assignments of partially deuterated Anabaena Sensory Rhodopsin, investigation of solvent exchangeability and deuterium isotope effects

MSc Thesis Presentation

Candidate: David Bolton

Abstract

Magic angle spinning solid-state NMR has been used with great success in studying membrane proteins in a native-like environment. These typically carbon-detected experiments are very time consuming, require a large amount of multiple samples with various levels of isotopic labeling. Through the use of proton detected experiments coupled with fast MAS we can reduce the total experimental time and number of required samples. Technological developments and labelling techniques (ultrafast MAS and isotopic dilution with 2H) have made proton detection in solid-state NMR a powerful tool. Membrane proteins however, remain a difficult subject of study. In this thesis, assignments of both exchangeable and non-exchangeable amide protons were made. With these assignments, solvent accessibility as well as deuterium isotope effects on chemical shifts were evaluated to help the assignment ant adjustment of assignment of proteins in future studies.

Examination Committee:

• Dr. Michele Oliver, Chair (Engineering)
• Dr. Vladimir Ladizhansky, Advisor (Physics)
• Dr. Leonid Brown, Co-Advisor (Physics)
• Dr. George Harauz, Examination Committee (Molecular and Cellular Biology)