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University of Groningen Biochemical and structural insights in bacterial B-type vitamin transporters of the Pnu family Singh, Rajkumar

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University of Groningen

Biochemical and structural insights in bacterial B-type vitamin transporters of the Pnu family Singh, Rajkumar

DOI:

10.33612/diss.109930154

IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below.

Document Version

Publisher's PDF, also known as Version of record

Publication date: 2020

Link to publication in University of Groningen/UMCG research database

Citation for published version (APA):

Singh, R. (2020). Biochemical and structural insights in bacterial B-type vitamin transporters of the Pnu family. University of Groningen. https://doi.org/10.33612/diss.109930154

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Biochemical and structural insights in

bacterial B-type vitamin transporters of the

Pnu family

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ISBN: 978-94-034-2293-0

ISBN: 978-94-034-2294-7 (electronic version)

Printed by: Gildeprint, Enschede

The research described in this thesis was carried out in Membrane Enzymology Department, Groningen Institute Biomolecular Science and Biotechnology, Faculty of Science and Engineering, Nijenborgh 4, 9747 AG, Groningen, The Netherlands (except chapter six). The research was financially supported by the Netherlands Organization for Scientific Research (NWO) and the European Research Council (ERC).

Cover design: Siddhartha Omar and Rajkumar Singh

Cover image: The thesis cover shows embedded membrane proteins of Pnu family PnuT, PnuX and PnuC in green, blue and pink respectively. These proteins transport their respectieve substrates as shown in the image.

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Biochemical and structural insights in

bacterial B-type vitamin transporters of

the Pnu family

PhD thesis

to obtain the degree of PhD at the University of Groningen

on the authority of the

Rector Magnificus Prof. C. Wijmenga and in accordance with

the decision by the College of Deans. This thesis will be defended in public on

Monday 6 January 2020 at 16.15 hours

by

Rajkumar Singh

born on 25 June 1983 in Mau, India

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Supervisor

Prof.D.J.Slotboom

Assessment Committee

Prof.B.Poolman Prof.A.J.M.Driessen Prof.V. Borshchevskiy

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TABLE OF CONTENTS

Chapter 1

Introduction 9

Chapter 2

PnuT uses a facilitated diffusion mechanism for thiamine uptake 41

Chapter 3

Crystallization screening of thiamin transporter-PnuT protein 61

Chapter 4

Structural and functional characterization of NadR from Lactococcus lactis 77

Chapter 5

A possible link between full length Pnu, SemiSWEET and SWEET transporters with preliminary study of Semi Pnu transporters 103

Chapter 6

Cryo-electron microscopic structure of SecA protein bound to the 70S ribosome 143

Summary

171

Samenvatting

173

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