University of Groningen
Insights into the transport mechanism of energy-coupling factor transporters
Stanek, Weronika Karolina
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Publication date: 2018
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Stanek, W. K. (2018). Insights into the transport mechanism of energy-coupling factor transporters. University of Groningen.
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Insights into the transport mechanism of
Energy-Coupling Factor transporters
Weronika Karolina Stanek
2018
Printed by: Ipskamp Printing
Cover photo: Roxana/ stock.adobe.com ISBN (printed version): 978-94-034-1052-4 ISBN (electronic version): 978-94-034-1051-7
The work described in this thesis was carried out in the Membrane Enzymology group of the Groningen Biomolecular Sciences and Biotechnology Institute of the University of Groningen, the Netherlands. The research was financially supported by the Netherlands Organization for Scientific Research (NWO) and the European Research Council (ERC). Copyright © 2018 by Weronika Stanek. All rights reserved. No parts of this thesis may be reproduced, stored in a retrieval system or transmitted in any form or by any means, without the permission of the author.
Insights into the transport mechanism of
Energy-Coupling Factor transporters
PhD thesis
to obtain the degree of PhD at the University of Groningen
on the authority of the Rector Magnificus Prof. E. Sterken
and in accordance with the decision by the College of Deans. This thesis will be defended in public on
Friday 26 October 2018 at 14:30 hours
by
Weronika Karolina Stanek
Promoters Prof. D.J. Slotboom Prof. B.Poolman Assessment committee Prof. M. Heinemann Prof. M.W. Fraaije Prof. F. MacMillan
Table of Contents
CHAPTER 1 General introduction to ECF-type ABC transporters 9
CHAPTER 2 Competition between different S-components for the shared Energy Coupling Factor module in Energy-Coupling Factor transporters 37
CHAPTER 3 In vitro reconstitution of dynamically interacting integral membrane subunits of Energy-Coupling Factor transporters 49
CHAPTER 4 Sample optimization for Single-molecule FRET analysis of S-component association with and dissociation from the ECF module 65
CHAPTER 5 Biochemical characterization of transport by type II Energy-Coupling transporters specific for pantothenate and folate 79
CHAPTER 6 Effect of lipids on the ECF transporters activity 107
CHAPTER 7 Three attempts to test the toppling hypothesis of S-components 119
CHAPTER 8 Optimization of expression constructs for heterologous production of ECF transporters 133
SUPPLEMENTARY TABLES 152
CONCLUSIONS AND OUTLOOK 162
NEDERLANDSE SAMENVATTING 166
STRESZCZENIE W JĘZYKU POLSKIM 169
