présentée l'Université Louis Pasteur Strasbourg I Pour le titre de

Sylvine Raverdy

Description

Niveau: Supérieur, Doctorat, Bac+8
THESE présentée à l'Université Louis Pasteur Strasbourg I Pour le titre de : Docteur de l'Université Louis Pasteur de Strasbourg Par Sylvine Raverdy Independent phosphoglycerate mutase and pyruvate phosphate dikinase: potential targets for drug discovery in filariasis Soutenue publiquement le 30 mai 2008 Devant les membres du jury Rapporteur Interne Dr. Michel Labouesse, Biologie cellulaire et développement, IGBMC, Strasbourg Rapporteurs externes Dr. Frédéric Bringaud, Microbiologie cellulaire et moléculaire et Pathogénicité, Bordeaux Dr. Kenneth Pfarr, Institute for Medical Microbiology, Immunology and Parasitology, Bonn Co-Directeurs de Thèse Prof. Jean-Marc Jeltsch, Phytopathologie, ESBS, Strasbourg Dr. Clotilde Carlow, Parasitology department, New England Biolabs, Ipswich

• acid abts


• card agglutination


• using genomic


• fructose-bisphosphate aldolase class


• coding deoxyribonucleic acid


• desoxyribonucleotid acid dpgam


• phosphoglycerate mutase


• adenosine diphosphate

Sujets

• Carlow
• Louis Pasteur University
• Kenneth

Informations

• Publié par: profil-zyak-2012
• Publié le: 01 mai 2008
• Nombre de lectures: 29
• Langue: English
• Poids de l'ouvrage: 4 Mo

Extrait

THESE
présentée à l’Université Louis Pasteur Strasbourg I
Pour le titre de :
Docteur de l’Université Louis Pasteur de Strasbourg
Par
Sylvine Raverdy
Independent phosphoglycerate mutase and
pyruvate phosphate dikinase: potential
targets for drug discovery in filariasis
Soutenue publiquement le 30 mai 2008
Devant les membres du jury
Rapporteur Interne
Dr. Michel Labouesse, Biologie cellulaire et développement, IGBMC, Strasbourg
Rapporteurs externes
Dr. Frédéric Bringaud, Microbiologie cellulaire et moléculaire et Pathogénicité,
Bordeaux
Dr. Kenneth Pfarr, Institute for Medical Microbiology, Immunology and
Parasitology, Bonn
Co-Directeurs de Thèse
Prof. Jean-Marc Jeltsch, Phytopathologie, ESBS, Strasbourg
Dr. Clotilde Carlow, Parasitology department, New England Biolabs, Ipswich Acknowledgements
I would like first of all to thank Dr. Bringaud, Dr. Labouesse and Dr. Pfarr for
accepting to be part of my committee and take the time of reading and commenting my
manuscript. Specialist’s opinion is always valued.
I would like to thank my two thesis advisors: Dr Clotilde Carlow and Prof. Jean-
Marc Jeltsch. Thanks Tilda for taking the chance of having a PhD student for the first
time, allowing me to develop in my project and to have the chance to be involve in such a
project. Thank you Prof. Jeltsch, you have always been there since the ESBS years and I
really appreciated your input in my work. I want to thanks all my colleagues from the
team, past and present especially Yinhua and Jeremy, you have been there since the
beginning and yours lab tricks and advices in and out of the lab have always been useful.
For the other members of the team, I’ll remember the weekly lab meeting where so many
ideas arise to resolve the everyday lab problems but also the informal lab meetings, in the
middle of the day because someone would have an idea. This team and all its members
were wonderful, helpful and always supportive.
I am very grateful to Dr. Donald Comb and Jim Ellard for allowing me to perform
my Ph.D studies in such a great environment and for their material support. I thank all
the NEB family. This company is a real paradise and a well of knowledge and sympathy.
Thanks for making feel at home. Thanks to all the persons who helped me one way or
another to write; correct and proofread this manuscript.
I thank all the friends I made along the way, Erica, dear I wish you all the
happiness and a great wedding, the French team, especially Sophie, Vanessa and Anne-
Lise, you three are great, and I hope to hear back from you soon! But I also want to
thank my new subie family with a special wink to Anita and Sarah, girls, you’ll see me
again. The Team Maplethropee, Mikey, Audrey, Dan, Lesley and Andy, thanks for
helping getting theses roles, all the good time spend together and the moral support you
gave me. Thanks to my roomies, Helen and Anulkah, it was really fun and great to live
with you. Good luck in the sunny land of California.
Last but not least, I want to thank my family who always encouraged me to go for
what I wanted. Well here I am. Table of contents
1Abbreviations…………………………………………………………………..
4General Introduction……………………………………………………….
5Neglected diseases and filarial parasites……………………………...........
Lymphatic filariasis…………………………………………………………………. 5
Onchocerciasis………………………………………………………………………. 9
12Treatment………………………………………………………………………….
Ivermectin…………………………………………………………………………… 12
Albendazole………………………………………………………………………….. 13
Diethylcarbamazine…………………………………………………………………. 14
Wolbachias as a drug target.………………………………………………………... 14
15Drug discovery process………………………………………………………...
16Using genomic and functional (C. elegans) data to identify new drug targets…….
Cofactor independent phosphoglycerate mutase as a potential drug target in
filarial nematodes and Wolbachia………………………………………………….. 18
Cofactor independent phosphoglycerate mutase as a drug target in African
trypanosomes ……………………………………………………………………….. 20
Genomic analysis of the glycolysis pathways in nematodes and Wolbachia……… 23
Pyruvate phosphate dikinase as a drug target in Wolbachia……………………… 25
Goals of the present study……...…………………………………………………… 26
28Results.....................................................................................................................
Chapter 1: Molecular and biochemical characterization of nematode cofactor
independent phosphoglycerate mutases……………………………………………. 29
Chapter 2: Expression of phosphoglycerate mutase from the Wolbachia
endosymbiont of Brugia malayi in the yeast Kluyveromyces lactis………………... 57
Chapter 3: Cofactor-independent phosphoglycerate mutase is an essential gene
in procyclic form Trypanosoma brucei……………………………………………... 75Table of contents
Chapter 4: The Wolbachia endosymbiont of Brugia malayi has an active
pyruvate phosphate dikinase……………………………………………………….. 93
Chapter 5: Identification of peptide ligands of independent phosphoglycerate
mutase using phage display libraries……………………………………………….. 110
Chapter 6: Functional characterization of the unrelated phosphoglycerate
mutases of Escherichia coli………………………………………………………… 129
153General Discussion and Conclusions………………………………...
160Materials and Methods…………………………………………………….
168Synthèse en Français………………………………………………………..
169Introduction.............................................................................................................
182Buts de l’étude………………………………………………………...…………
183Discussions et Conclusions…………………………………………………….
190References……………………………………………………………………….Abbreviations
2-PG: 2-phosphoglycerate
3-PG: 3-phosphoglycerate
aa: amino-acid
ABTS: 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)
ABZ: Albendazole
ADP: Adenosine Diphosphate
AMP: Adenosine Monophosphate
ATP: Adenosine Triphosphate
BSA: Bovine serum albumine
CATT: card agglutination test for trypanosomes
cDNA: coding deoxyribonucleic acid
CIATT: Card Indirect Agg