University of Groningen
Structural and biochemical characterization of Roco proteins Terheyden, Susanne
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Publication date: 2018
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Terheyden, S. (2018). Structural and biochemical characterization of Roco proteins. University of Groningen.
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Chapter 1
Introduction
The unconventional G-protein cycle of LRRK2 and Roco proteins
Susanne Terheyden, Laura M. Nederveen-Schippers and Arjan Kortholt
This chapter has been published in: Soc. Trans. 2016, 44, 1611–1616.
Aim of the Thesis
Roco proteins, including LRRK2, in general are very complex proteins in numerous different aspects. Not only have they been linked to nearly every process within the cell, but also their domain architecture and self-regulation is complicated and not yet completely understood. It gave the impression of a large puzzle where now and then pieces were discovered but they didn’t seem to easily fit to each other yet. This thesis aims to contribute a piece of the puzzle by shedding light on the activation mechanism of the Roco protein family’s name giving feature, the RocCOR domain tandem. Since several of the PD mutations are located in these two domains, it seems clear that the RocCOR domain tandem has important functions in LRRK2 and Roco proteins. Independent of each other, several groups have presented indication that the Roc domain is not a classical small G-protein but might act differently in Roco proteins, but also that the Roc domain plays a central role in LRRK2 activation and PD pathogenesis. Therefore the aim of my thesis is to investigate and biochemically characterize the RocCOR domain tandem and thereby contribute to the understanding of Roco proteins, especially LRRK2 and its malfunction in PD.
The second chapter describes the structure and biochemical features of the RocCOR domain tandem lacking the C-terminal dimerization domain. In the third chapter we discovered that the Chlorobium tepidum (Ct) Roco protein has the ability to monomerize upon GTP binding which was actually very surprising and at first sight opposed to our initial hypothesis, that Roco proteins belong to the group of GADs that dimerize in the presence of GTP. In the fourth chapter we set out to biochemically characterize this Ct Roco protein and several other prokaryotic Roco proteins thoroughly to further elucidate the kinetic behaviour of the hydrolysis reaction. Chapter 5 presents the structures of the Methanosarcina barkeri (Mb) RocCOR tandem bound to different nucleotides. Finally chapter 6 summarizes all these findings in the context of LRRK2, puts some puzzle pieces together to form the bigger picture.
