Universiteit Leiden

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Proefschrift

Applications of paramagnetic NMR spectroscopy for protein research

The aim of the research presented in this thesis was to develop new methods forchallenging systems in liquid-state NMR using paramagnetic effects generated by thetwo-armed probe CLaNP-5.

Auteur
Lescanne, M.
Datum
29 mei 2018
Links
Thesis in Leiden Repository

The aim of the research presented in this thesis was to develop new methods forchallenging systems in liquid-state NMR using paramagnetic effects generated by thetwo-armed probe CLaNP-5. Chapter 1 is an introduction about NMR assignmentdevelopment and challenges and a brief opening to the theory of paramagnetic effectsand their applications. Chapter 2 describes the upgrade of PARAssign software and itsability to assign and assess methyl groups of a 25 kDa protein using experimentalpseudocontact shifts from 1 to 3 paramagnetic centers. Chapter 3 describes theimplementation of residual dipolar coupling into PARAssign software as extra datasetinput in addition to the pseudocontact shifts. Applications are given for residueselective isotope labeled protein. Chapter 4 depicts the use of pseudocontact shiftsused to probe small methyl group movements and re-orientation under ligand binding.Chapter 5 gives some clues about how paramagnetic effect such as RDC but also PCScan provide information for challenging highly dynamic system. Chapter 6 givesinsights for PCS application to very large multimeric protein, the challenges andlimitations are discussed. Chapter 7 provides a discussion and few perspectives aboutthe work carried out and presented in this thesis.

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