Feeling sugar-protein interactions using carbon nanotubes : a molecular
recongition force microscopy study
Klein, D.C.G.
Citation
Klein, D. C. G. (2004, November 11). Feeling sugar-protein interactions using carbon
nanotubes : a molecular recongition force microscopy study. Retrieved from
https://hdl.handle.net/1887/106077
Version:
Publisher's Version
License:
Licence agreement concerning inclusion of doctoral thesis in the
Institutional Repository of the University of Leiden
Downloaded from:
https://hdl.handle.net/1887/106077
Cover Page
The handle http://hdl.handle.net/1887/106077 holds various files of this Leiden University
dissertation.
Author:
Klein, D.C.G.
Feeling sugar-protein interactions
using carbon nanotubes
A digital version of this thesis can be downloaded from http://www.physics.leidenuniv.nl
Cover:
Front: Artist impression of a functionalized carbon nanotube AFM tip
probing a surface that contains a receptor. Both sample surface and tip are immersed in liquid. The drawing was made by Henriette Jensenius.
Back: from left upper corner to right lower corner:
AFM image taken in liquid of a Chinese hamster ovary cell, picture was taken by Maarten van Es; molecular structure of mannose with a three-carbon linker and an amine group at the end, image was made by Karin Sliedregt-Bol; AFM image of purple membrane in liquid, trimers of bacteriorhodopsin are arranged in a hexagonal lattice;
Pisum sativum lectin immobilized on mica, imaged by AFM in liquid,
force-distance curve taken with a carbon nanotube tip on a carboxyl-SAM in liquid, functionalization cell for AFM tips.
Background: modified mica surface imaged by light microscopy in air.
Feeling sugar-protein interactions
using carbon nanotubes
A molecular recognition
force microscopy study
PROEFSCHRIFT
ter verkrijging van
de graad van Doctor aan de Universiteit Leiden, op gezag van de Rector Magnificus Dr. D.D. Breimer,
hoogleraar in de faculteit der Wiskunde en Natuurwetenschappen en die der Geneeskunde,
volgens besluit van het College voor Promoties te verdedigen op donderdag 11 november 2004
klokke 14.15 uur door
Dionne Clara Gertrud Klein
PROMOTIECOMMISSIE
Promotores: Prof. dr. J.W.M. Frenken
Prof. dr. J.W. Kijne
Co-promotor: Dr. T.H. Oosterkamp Referent: Dr. P. Hinterdorfer
(University of Linz, Austria) Overige leden: Dr. Th. J. Aartsma
Prof. dr. P.H. Kes
Dr. R. McKendry
(University College London, UK) Prof. dr F.W. Saris
Prof. dr. H.P. Spaink
Contents iii
Contents
Chapter
Page
1 Introduction 1
1.1 Introduction and motivation 2
1.2 Receptor-ligand interactions 4
Historical background of AFM in receptor-ligand research 5 Sugar-lectin interactions in this thesis 5
1.3 Atomic force microscopy 6
Molecular recognition force microscopy 7
1.4 Carbon nanotubes 8
1.5 Outline of this thesis 10
2 An AFM study of FIN filaments: 13 filaments in between the nuclei of dividing
Saccharomyces cerevisiae (yeast) cells
2.1 Introduction 14
Yeast cell division 14
Microtubuli 15
Fin1 protein 16
Confocal fluorescence microscopy 16
Electron microscopy 17
Goals 17
2.2 Methods 17
Fin 1 preparation 17
Sample preparation 18
Atomic force microscopy 18
Control experiments 19
2.3 Discussion and conclusion 20
3 Imaging biological molecules and membranes under 23 physiological conditions
3.1 Introduction 24
3.2 Contact mode in liquid 24
Native purple membrane 27
Experimental details 28
Light harvesting 2 complex (LH2) 30
Contents iv
Acoustic driving 32
Pea lectin dimers 33
MAC mode: magnetic driving 34
Chinese hamster ovary (CHO) cells 34
3.4 Conclusions 36
4 Covalent immobilization of single proteins on mica for 39 molecular recognition force microscopy
4.1 Introduction 40
4.2 Experimental procedures 42
Surface modification 43
Protein immobilization 43
AFM imaging 44
4.3 Results and discussion 45
4.4 Conclusions 49
5 Carbon nanotubes as nanometer-sized probes for AFM 51
5.1 Introduction 52
5.2 Single-walled carbon nanotube AFM tips 52 Production: chemical vapor deposition 52
Mounting procedure 54
Intermezzo: carbon nanotube mechanics 56
Nanotube shortening 59
Making the tip water-proof 60
5.3 Imaging antibody molecules with SWNT AFM tips 61 5.4 Multi-walled carbon nanotube AFM tips 64
Production: arc discharge 64
Mounting procedure 65
Making the tip water-proof 65
Nanotube shortening 66
5.5 High-resolution TEM results 67
Open versus closed nanotubes 67
5.6 Conclusions 69
6 Carbon nanotube functionalization for high-resolution 71 molecular recognition force microscopy
6.1 Lateral resolution in molecular recognition force 72 microscopy
6.2 Chemical functionalization of AFM tips for 72 high-resolution MRFM
6.3 Force titration measurements 74
Carboxylic acid both on tip and surface 75 Carboxylic acid on the tip, hydroxyl terminated surface 78
Contents v
Experimental procedures 80
6.6 Conclusions 81
7 Molecular recognition force microscopy on pea lectin 83 and mannan binding lectin 7.1 Introduction 84
7.2 Interpretation of phase images; qualitative and 85
quantitative aspects MRFM with short spacers: how to detect unbinding 88 events 7.3 Pisum sativum lectin 88
Experimental details 89
In air 89
In liquid 90 MRFM with short spacers: how to detect unbinding 92 events, part II 7.4 Mannan binding lectin 95
Experimental details 96
In air 96
Preliminary experiments in liquid 97
7.5 Conclusions 102
8 Summarizing discussion and outlook 105
8.1 Summarizing discussion 106
8.2 Technical outlook 106
8.3 Scientific challenges 107
Appendices
Carbon nanotube mechanics Gold crystal preparation
Acknowledgements Summary
Glossary
Samenvatting voor de leek Curriculum vitae
