Investigation of the ubiquitin-specific protease UBP41 and of the lysosomal cysteine proteases cathepsin-L and cathepsin-B as potential mediators of proapoptotic signalling [Elektronische Ressource] / submitted by Andreas Gewies

Gewies, Andreas

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• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2003
• Nombre de lectures: 65
• Langue: English
• Poids de l'ouvrage: 3 Mo

Extrait

Fakultät für Biologie der Ludwig-Maximilians-Universität
München





DISSERTATION





Investigation of the ubiquitin-specific protease UBP41 and of
the lysosomal cysteine proteases cathepsin-L and cathepsin-B
as potential mediators of proapoptotic signalling








Submitted by

Andreas Gewies







Tag der Einreichung: 10. November 2003
Day of Submission:

Erstgutachter: PD Dr. Stefan Grimm
First Referent:

Zweitgutachter: Prof. Dr. Charles David
Second Referent:

Begutachter/Accessors:
Prof. Dr. Walter Schartau,
PD Dr. Angelika Böttger,
Prof. Dr. Michael Boshart,
Prof. Dr. Jürgen Soll

__________________________________________________


Tag der mündlichen Prüfung: 20. Januar 2004
Day of Viva Voce:

Prüfer / Examiners:
PD Dr. Stefan Grimm,
Prof. Dr. Charles David,
Prof. Dr. Walter Schartau,
PD Dr. Angelika Böttger Short INDEX
Short INDEX
PhD candidate declaration ...................................................................................... ii
Dedication .............................................................................................................. iii
Acknowledgement .................................................................................................. iv
ApoPainting.............................................................................................................v

Long INDEX ................................................................................................... vi

SUMMARY ...........................................................................................1


1 INTRODUCTION .........................................................................5

1.1 APOPTOSIS..............................................................................................5
1.2 APOPTOSIS AND THE UBIQUITIN/PROTEASOME SYSTEM ...........................26
1.3 LYSOSOMES AND THEIR POSSIBLE INVOLVEMENT IN APOPTOSIS.................36
1.4 SPECIFIC AIMS OF THIS THESIS...............................................................39

2 MATERIALS AND METHODS ..................................................40

2.1 MATERIALS............................................................................................40
2.2 METHODS..............................................................................................46

3 RESULTS ..................................................................................61

3.1 IDENTIFICATION AND CHARACTERIZATION OF UBP41 AS A
DOMINANT APOPTOSIS-INDUCING FACTOR...............................................61
3.2 INVESTIGATION OF A POSSIBLE INVOLVEMENT OF CATHEPSIN-L AND
CATHEPSIN-B IN APOPTOSIS INDUCTION ..................................................82

4 DISCUSSION.............................................................................93

4.1 UBP41 - A DOMINANT APOPTOSIS-INDUCING FACTOR ...............................94
4.2 CATHEPSINS AND THEIR POSSIBLE INVOLVEMENT IN PROAPOPTOTIC
SIGNALLING .........................................................................................111

References..........................................................................................................116
Publications.........................................................................................................136
Curriculum Vitae..................................................................................................138
Abbreviations.......................................................................................................139
i
PhD candidate declaration

PhD candidate declaration

I certify that the thesis entitled

“Investigation of the ubiquitin-specific protease UBP41 and of the lysosomal cysteine proteases
cathepsin-L and cathepsin-B as potential mediators of proapoptotic signalling”,

submitted for the degree of

Dr. rer. nat.,

is the result of my own, independent work, that all of the following material was written by myself,
and that I did not use any other sources of information than those indicated by the given references.


Erklärung

Hiermit versichere ich, dass die vorliegende Promotionsarbeit das Ergebnis meiner eigenständigen
Arbeit ist und dass ich dieses Manuskript persönlich verfasst habe. Zudem habe ich keine anderen als
die angegebenen Quellen und Hilfsmittel verwendet.









München, 10. November 2003 Andreas Gewies
ii
Dedication








I like to dedicate this work to my family,
to my parents Magdalena and Norbert Gewies,
to my sisters Christiane and Gabriela
as well as to my little brother Marcel.

I am looking back on those former times
that shaped me to what I am today,
that helped me to develop my various interests
and that gave me the opportunity to receive a decent education.

A special dedication to my beloved fiancée Bianca
who has accompanied me during the last two years of this work
and who means to me a wonderful world beyond science.

In memory of my grandfather Wilhelm Riedel,
who to me will always be an example of
modesty and decency.

iiiAcknowledgement
Acknowledgement

The here presented doctoral thesis is based on the data and results obtained during my work at the
Max-Planck-Institute for Biochemistry in Martinsried, Germany, from January 2000 until August
2003. Supervisor was PD Dr. Stefan Grimm as the principal investigator of the independent Junior
Research Group with focus on “Programmed Cell Death”. This work was supported by the
Sonderforschungsbereich 469, by the Bavarian Government, Xantos Biomedicine AG, and Roche
Diagnostics.

_________________________________________________________________________________

First of all, I would like to express my appreciation and thanks to my supervisor PD Dr. Stefan Grimm
for giving me the opportunity to become a member of his group, for his support and his critical
reviews of my work, by this guiding me to the completion of this thesis as well as to the successful
publication of its results.

I also wish to thank Professor Dr. Charles David as second supervisor for his constant support of the
PhD students of our group, including myself, and his intense interest in our research. Thanks also to all
other referees of this thesis work.

My special thanks to my colleagues and friends in the laboratory “Programmed Cell Death” for their
stimulating comments, interesting discussions, technical support and especially for creating a pleasant
working environment. Thanks to Timur Albayrak, Manuel Bauer, Sandra Bernard, Erik Braziulis, Ulla
Cramer, Thomas Mund, Jasmina Putnik, Volker Scherhammer, Nicole Schönfeld, Alexis Schubert,
Frank Voß, but also to colleagues from other groups. I will miss you and at the same time hope to keep
in touch!

Finally, I like to thank my former colleagues from the Department of Pathology at the University of
Iowa in Iowa City, USA, especially Prof. Michael B. Cohen M.D. and Dr. Oskar W. Rokhlin for
introducing me to the field of apoptosis research and for giving me the opportunity to independently
practice the scientific research and writing process.

iv
Apoptosis –
the end of an individual cell, at the same time a martyrdom in favor of the organism
as a whole. A well-ordered process, genetically determined, also called “programmed
cell death” what sounds somewhat artificial. However, apoptosis is as natural as the
falling of leaves from a tree in autumn.
vLong INDEX
Long INDEX


Short INDEX.............................................................................................................i
PhD candidate declaration ...................................................................................... ii
Dedication .............................................................................................................. iii
Acknowledgement .................................................................................................. iv
ApoPaintingv
Long INDEX ........................................................................................................... vi



SUMMARY .................................................................................... 1



1 INTRODUCTION................................................................... 5

1.1 APOPTOSIS ..........................................................................................5
1.1.1 The development of the term apoptosis................................................5
1.1.2 The significance of apoptosis................................................................5
1.1.3 Morphological fea