Modeling cell adhesion and its control mechanisms with a vesicle-substrate system [Elektronische Ressource] / Ana-Sunčana Smith

Asmith

Sujets

• Biologie

Informations

• Publié par: technische_universitat_munchen
• Publié le: 01 janvier 2005
• Nombre de lectures: 22
• Langue: English
• Poids de l'ouvrage: 23 Mo

Extrait

PHYSIK-DEPARTMENT





Modeling cell adhesion and its control
mechanisms with a vesicle-substrate
system

Dissertation
von

Ana-Sun čana Smith





TECHNISCHE UNIVERSITÄT
MÜNCHEN
Physik-Department
Der Technischen Universität München
Lehrstuhl für Biophysik




Modeling cell adhesion and its control mechanisms with a vesicle-
substrate system




Ana-Sun čana Smith




Vollständiger Abdruck der von der Fakultät für Physik der Technischen Universität
München zur Erlangung des akademischen Grades eines

Doktors der Naturwissenschaften (Dr. rer. nat.)

genehmigten Dissertation.



Vorsitzender:

Univ.-Prof. Dr. R. Netz



Prüfer der Dissertation:

1. Univ.-Prof. Dr. E. Sackmann em.
2. Univ.-Prof. Dr. U. Seifert, Universität Stuttgart





Die Dissertation wurde am 22.11.2004. bei der Technischen Universität München
eingereicht und durch die Fakultät für Physik am 16.12.2004. angenommen.










k ćerima i majkama v
Acknowledgments

I extend my thanks to all the people who helped me in my professional, personal and any
other life during my Doctorate degree. Many, however, warrant special mention:
Firstly, I would like to extend my sincerest thanks to my Doktorvater, Professor Erich
Sackmann for the unconditional trust he has placed in me. His contagious passion for
Biological Physics, great ideas, and the beautiful working environment that he created in
E22, were my driving forces. His ability to recognize and provide the support and
understanding which I needed, when I needed, has left me with a feeling of being an
extremely well-managed student. Danke für alles.
I am very lucky to have had the privilege to work with Professor Udo Seifert. Although
staying behind the scenes, he provided me with unrestrained supervision, and introduced
me to the field of theoretical biophysics. His sound advice and always immediate
response to my constant questions are greatly appreciated. Thank you for taking me into
your fold.
Dr. Barbara Lorz and Dr. Stefanie Gönnenwein were there when I just started. Their
experiments were both the inspiration and the grounding for my beginnings in the field of
vesicle adhesion. Equally unforgettable were the little after-hours chats and the coffee
breaks which made my days much warmer.
Thanks are extended to past and present members of E22, for the collegial atmosphere,
assistance and laughter which they provided in the day-to-day experience. Moments with
Dr. Kheya Sengupta and Dr. Gjertrud Maurstad will always be in my memories. I would
also like to mention Dr. Wolfgang Feneberg, Dr. Doris Heinrich and Dr. Felix Linke.
I am extremely grateful to the Frauenbeauftragte der Technische Universität München for
enabling me to participate in Hochschul- und Wissenschaftsprogramm (HWP II) that
awarded me a bursary for postgraduate students in the final stages of a Doctorate degree.
Particular words are also addressed to Christian Daniel, Katrin Prechtel, Jörg Uhde and
their partners Tina, Schüssel and Sabine for being much more than colleges - for being vi
friends. Thank you for opening the door to the German culture and helping with the
sometimes bewildering aspects of a life as a foreigner. To this end, my special thanks are
extended to the Oberschleissheimer Families Me đugorac and Peja čevi ć and also to Frau
Christine Singer for giving me and my family the impression of being welcomed and
home.
Somewhat strange paths took me to Australia for five months during my Doctorate. The
welcome extended to me by the entire Department of Applied Mathematics at the
Australian National University, Canberra, ACT is greatly appreciated. Especially, Dr.
Tim Senden, Dr. Gerd Schröder, Dr. Anke Karlsson with her partner Dr. Simon Bennett,
and Mr Holger Averdunk helped to make this visit a pleasant experience.
In the final months of my Doctorate, I spent extensive periods of time at the Institute for
Physics of the University of Zagreb, Croatia. Assoc. Professor Katarina Uzelac and her
Group provided support during this stage and for this I am very grateful. Dr. Davorin
Lovri ć, and the other members of the institute were a source of great cheering and fun.
With warmth in my heart, I have to thank my mother Solange Bariši ć and my sister Ivana
Tanovitski as well as all other members of my family for helping unconditionally and
decreasing the load whenever possible. Wherever you are, in Croatia, Russia,
Switzerland, Australia or France, you are in my thoughts.
At last, but as great as to the first, much praise must go to David and Lana: The sacrifices
you made enabled me to come to the point where I can write these final words. For this
and for the life you are sharing with me, thank you. vii
Table of Contents
ABSTRACT ix
ZUSAMMENFASSUNG xi
CHAPTER 1: PHYSICAL ASPECTS OF CELL ADHESION 2
1.1 THE CELL 4
1.1.1 CELL MEMBRANE STRUCTURE 5
1.2 CELL ADHESION 10
1.2.1 CONTROL MECHANISMS 11
1.3 MODEL SYSTEMS 12
1.3.1 ADHESION OF FLAT MEMBRANES 13
1.3.2 MEMBRANES IN THREE DIMENSIONS 18
1.4 REFERENCES 24
CHAPTER 2: MODELING VESICLE ADHESION 31
2.1 NOTATION 32
2.2 THE FREE ENERGY 34
2.3 MINIMIZING THE FREE ENERGY 38
2.3.1 MINIMIZATION WITH RESPECT TO THE CONSTITUENT VARIABLES 38
2.3.2 MINIMIZATION WITH RESPECT TO THE SIZE OF THE CONTACT ZONE 39
2.4 ALLOCATION OF LIGANDS IN THE CONTACT ZONE 41
2.4.1 LIMITING BEHAVIORS OF THE ALLOCATION FUNCTIONS
2.4.2 OVERALL BEHAVIOR OF THE ALLOCATION FUNCTIONS 42
2.4.3 BEHAVIORAL REGIMES OF THE ALLOCATION FUNCTIONS OF BOUND LIGANDS 43
2.5 THE EFFECTIVE ADHESION STRENGTH 53
2.5.1 LIMITING BEHAVIOR AND THE DEPENDENCE ON THE BINDING STRENGTH OF THE
LIGAND-RECEPTOR PAIR 54
2.6 CONSEQUENCES OF THE MODEL 58
2.7 REFERENCES 62
CHAPTER 3: ANTAGONIST-INDUCED UNBINDING 64
3.1 OBSERVATION OF ANTIBODY INHIBITION IN THE MODEL SYSTEM 66
3.1.1 ADHESION OF VESICLES TO THE SUBSTRATE 66
3.1.2 ANTAGONIST-INDUCED UNBINDING 67
3.2 QUANTIFICATION OF THE OBSERVED UNBINDING 73
3.2.1 THE LATERAL-PRESSURE MECHANISM - PHASE I 74
3.2.2 THE COMPETITIVE BINDING MECHANISM –PHASE II 81
3.3 IMPLICATIONS OF THE UNBINDING MECHANISMS 86
3.4 REFERENCES 89
viii
CHAPTER 4: FORCE AS A CONTROL MECHANISM 91
4.1 MODELING THE EXERTION OF FORCE 94
4.2 STRONG ADHESION 97
4.3 WEAK ADHESION 100
4.3.1 EXPERIMENTAL OBSERVATIONS – SHAPE DECOUPLING 100
4.3.2 EXPE – EVENTS IN THE CONTACT ZONE 103
4.4 REFERENCES 107
SUMMARY AND CONCLUSIONS 109
APPENDIX A: BENDING ENERGY OF A VESICLE SHAPE 114
A.1 DETERMINATION OF THE FITTING FUNCTION 114
A.2 DEPENDENCE OF THE COEFFICIENTS ON THE REDUCED VOLUME 119
A.3 REFERENCES 119
APPENDIX B: SPECIFICATIONS OF THE EXPERIMENTS 120
B.1 MATERIALS 120
B.2 METHODS 122
B.3 REFERENCES 125
APPENDIX C: CONTINUOUS MODEL FOR UNBINDING: STRONG ADHESION 126
C.1 PULLING THETHERS FROM ADHERED VESICLES 126
C.2 METHODOLOGY 130
APPENDIX D: CONTINUOUS MODEL FOR UNBINDING: WEAK ADHESION 137
D.1 EFFECTS OF A PULLING FORCE ON THE SHAPE OF A BOUND VESICLE 137
D.2 METHODOLOGY 144

BIBLIOGRAPHY 153

CURRICULUM VITAE 166

LIST OF PUBLICATIONS 167

LIST OF SYMBOLS AND ABBREVIATIONS 168
ix
Abstract
In order to study the process of cell adhesion and its underlying mechanisms, a
theoretical model system has been developed. The model consists of a functionalized
vesicle capable of interacting with an activated substrate. More specifically, mobile
ligands incorporated into the vesicle membrane can undergo specific interactions with
receptors immobilized on the surface of the substrate. Under suitable conditions, a zone
of contact is formed between the vesicle and the substrate, and a process known as
adhesion ensues.
The model is simply constructed yet conceptually rich. In particular, explicit
consideration is given to the following factors: (i) the enthalpy of binding (ii) the mobility
of the ligands through a contribution to the mixing entropy, (iii) the bending energy of the
vesicle shape, (iv) the finite number of ligands contained in the vesicle, and (v) the
constant density of receptors on the substrate. The equilibrium conditions of such a
canonical ensemble is studied in detail and several adhesion regimes, dependent upon the
densities of the system constituents and the binding strength of the ligand-receptor pair,
are identified.
Of the many interesting outcomes emerging from the model, the most important can be
hierarchically laid out as follows: (i) The explicit treatment of the bending energy shows
that the determination of the vesicle shape can be decoupled from the equilibration of
ligand-receptor binding in the contact zone. (ii) The density of bonds formed is found